Oral Care Compositions With Anticaries Activity

ABSTRACT

Oral care compositions with two or more components with a subtherapeutic anticaries effect, but, in combination, provide a therapeutic anticaries effect. Oral care compositions that are fluoride-free or lower in fluoride content, but have an equivalent or greater anticaries benefit than a composition comprising a therapeutic amount of fluoride ions.

FIELD OF THE INVENTION

The present invention is directed to compositions with anticariesactivity. The present invention is also directed to compositions withtwo or more anticaries agents that are individually subtherapeutic, butthat have an overall therapeutic anticaries activity. The presentinvention is also directed to fluoride-free compositions with anequivalent or greater anticaries benefit than a composition comprising atherapeutic amount of fluoride ions. The present invention is alsodirected to compositions comprising a therapeutic amount of fluoride andanother anticaries agent with a greater anticaries benefit than acomposition comprising a therapeutic amount of fluoride ions as the soleanticaries agent.

BACKGROUND OF THE INVENTION

The current consumer goods marketplace reflects an increasing awarenessof the entire lifecycle of a product, including the provenance of thevarious ingredients, packaging, and research methods used tosubstantiate a product's effectiveness. Consumers are rejectingpetrochemically derived ingredients and shifting the marketplace toresponsible-sourced and naturally-derived raw materials, recyclablepackaging, and minimally processed materials. Coupled with this trend isa rejection of fluoride for concerns, real or imagined, of its toxicityin drinking water, toothpaste, or both. This trend has provably led toincreases in cavities in consumers whom reject fluoride because thereare no viable alternatives to fluoride in over-the-counter oral careproducts for the prevention of cavities. Consequently, the currentmarketplace requires that consumers trade clean (giving up fluoride) foreffective (anticavity toothpaste).

As such, there is a need in the art for a fluoride-free, anticavitytoothpaste for consumers who reject fluoride in drinking water and/ortoothpaste. Additionally, there is a need in the art for the enhancementfor fluoride's effectiveness in a fluoride-containing toothpaste forconsumers that reject fluoride in the drinking water, but still use atherapeutically effective level of fluoride in an anticavity toothpaste.There is also a need in the art for the enhancement of fluoride'sactivity in a situation where fluoride is formulated at a subtherapeuticlevel in order to reduce overall fluoride exposure, which is desirablesituation for young children using anticavity toothpaste. There is alsoa need in the art to deliver the above compositions using naturallyderived ingredients whose efficacy is traditionally significantlydeficient that of traditional therapeutic levels of fluoride (i.e., 1100ppm F as NaF in a silica-based toothpaste).

SUMMARY OF THE INVENTION

This summary is provided to introduce a selection of concepts in asimplified form that are further described below in the detaileddescription. This summary is not intended to identify required oressential features of the claimed subject matter. Nor is this summaryintended to be used to limit the scope of the claimed subject matter.

Disclosed herein is an anticavity, fluoride-free oral care composition,wherein the composition has a rat caries score of about 60% or more of arat caries score of a positive control oral care composition, thepositive oral care composition comprising 1100 ppm of sodium fluoride.

Also disclosed herein is an oral care composition comprising (a) a firstsubtherapeutic anticaries agent (b) a second subtherapeutic anticariesagent, wherein the oral care composition is free of a fluoride sourceand the first and second subtherapeutic anticavity agents collectivelyhave a therapeutic anticavity benefit.

Also disclosed herein is an oral care composition comprising (a) ananticaries drug composition comprising fluoride; and (b) asubtherapeutic anticaries agent, wherein the subtherapeutic anticariesagent is free of fluoride, wherein the oral care composition has atherapeutic benefit greater than the anticaries drug composition.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows rat caries results for the invention relative to thefluoride control products.

FIG. 2 shows iPGRM dose response of chlorhexidine (% acid reduction vCrest Cavity Protection) along with the iPGRM performance of the novelcompositions

FIG. 3 shows iPUM behavior of different calcium sources (ratio ofbiofilm calcium content following treatment relative to the biofilmcalcium content following treatment with Crest Cavity Protection) alongwith the iPUM performance of the novel compositions

FIG. 4 shows F-Free HAP solubility reduction with respect to Sn contentalong with the F-Free HAP Solubility reduction of typical Sn/silicatoothpaste compositions.

FIG. 5 shows actual v predicted rat caries percent reduction vs. placebo(left) and the plot of residuals (right).

DETAILED DESCRIPTION OF THE INVENTION

Caries is a process by which tooth damage occurs by exposure to plaqueacids and the subsequent net demineralization of the tooth. Bacteria inthe plaque produce organic acids from the metabolism of fermentablecarbohydrates to which they have access when a person eats.Consequently, avenues for intervention in caries generally involve thesuppression of acid formation and/or the stabilization of the toothsurface. In the present invention, we have considered four mechanismsthrough which this may occur, namely: i) suppressing acid formation viaantibacterial action; ii) reducing enamel solubility through a calciumco-ion effect; iii) reducing enamel solubility through a fluoride co-ioneffect; and iv) reducing enamel solubility through surface adsorbedstabilizers. These mechanisms are considered essential for the reductionin human cavities. Until now, the one favored by nearly everyanti-cavity product on the market is exclusively the reduction of enamelsolubility through a fluoride co-ion effect.

Thus, the present invention is directed at compositions that can treatcaries and prevent cavities comprising two or more subtherapeuticanticaries agents, such as an antibacterial agent, a surface-adsorbedenamel stabilizing agent, a calcium ion source, and/or a fluoride ionsource. Additionally, the present invention is directed at compositionswith improved anticaries activity comprising a therapeutic amount offluoride ions and one or more subtherapeutic anticaries agents, such asan antibacterial agent, a surface-adsorbed enamel stabilizing agent, orcalcium ion source.

Additionally, the present invention is directed to creating atherapeutic oral care composition that is a combination of two or moresub-therapeutic compositions using a model for the performance of oralcare compositions generally in rat caries along the vectors of fluorideco-ion effect, calcium co-ion effect, Sn-free antibacterial efficacy,and F-Free hydroxyapatite solubility reduction via surface adsorption.Such a model will allow the rapid, ethical, and responsibleidentification of ingredients that minimizes or virtually eliminatesanimal testing for new therapeutic anti-caries oral care compositions tocreate fluoride-free and enhanced-fluoride compositions.

Since the discovery of fluoride there have been numerous attempts in theart to find alternatives to, or enhancements for, fluoride. Thoseattempts have generally favored one of the four mechanisms listed above,most especially plaque acid suppression. The disclosed methods andcompositions of the present application are radically different fromwhat was presented in the art. While not wishing to be bound by theory,it is believed that subtherapeutic amounts of compositions suppliedsimultaneously would deliver, in total, a composition capable ofreducing cavities. Thus, the present invention is directed tocompositions comprising at least two subtherapeutic compositions inaccord with the mechanisms described above, yet the disclosedcompositions delivered a reduction in caries as measured in the ratcaries model at least equivalent to 1100 ppm F as sodium fluoride andsuperior to that of the 1100 ppm F control when fluoride was included,as shown in FIG. 1. Additionally, the disclosed compositions led to areduction of caries in the rat sulcus and interproximal spaces thatexceeded the effectiveness of fluoride. This unexpected resultdemonstrated that this approach was novel as nowhere in the art was ittaught that a combination of subtherapeutic compositions could beeffective for the prevention of cavities.

The rat caries results, presented in FIG. 1, indicated that Ex 1breduced caries relative to sodium fluoride/silica andmonofluorophosphate/silica compositions in a placebo-controlled study.Ex. 1b which did not have fluoride, had a total caries reductionequivalent to the sodium fluoride/silica control product; however, ithad an unexpectedly large reduction in sulcal caries relative tocontrol. The disclosed compositions were designed to work on the toothand in the biofilm with a collection of subtherapeutic compositions.This has led to a remarkable reduction in sulcal and interproximalcaries with respect to the control compositions. When fluoride is addedto the disclosed compositions, as in Ex. 1d, the reduction in caries isenhanced with a notable reduction in smooth surface caries.

It was unexpectedly found that the non-fluoride mechanisms can have alarge contribution to the reduction in caries considering that such anapproach had not be disclosed in the art. In order to better understandthis result, a set of laboratory tests were performed to measure thecontribution of each subtherapeutic composition to the compositiontested in the rat. Individual methods have been developed to measure theefficacy of compositions along the above described intervention vectors.These methods are the: i) Sn-Free in vitro plaque glycolysis andregrowth method (Sn-Free iPGRM); ii) in vitro plaque uptake method forcalcium (iPUM-Ca); iii) F-Free hydroxyapatite solubility reductionmethod (F-Free HAP); and iv) ADA one-minute fluoride release (ADA). Aperson of ordinary skill in the art would recognize that someingredients, such as Sn, have both an antibacterial effect and a HAPsurface stabilization effect. Such behavior complicates the analysis ofa composition's performance; therefore, the entire contribution of suchingredients is considered through a single mechanism only. Thus, for thepurposes of this invention, a composition's antibacterial efficacyshould be determined with respect to its Sn placebo or by otherexperimental design approaches that correctly account single variablyfor the contribution of Sn in the iPGRM. Similarly, for the purposes ofthis invention, a composition's ability to reduce hydroxyapatitesolubility should be determined using the fluoride-free version of thecomposition (if it contains fluoride) or by some other experimentaldesign that correctly controls for fluoride's contribution. TABLE 2illustrated the results of the characterization of the novelcompositions illustrated in TABLE 1 and control toothpastes using thefour different methods indicated above with their corresponding ratcaries scores, as disclosed in the Example section.

During the discovery of fluoride, several rat caries tests were runexploring both the effectiveness of fluoride as well as alternatives.The catalogue of rat caries experiments were meta-analyzed to illustratethe single variable behavior of the previously mentioned interventionvectors. As will be illustrated later, this let us evaluate the efficacyof different vectors relative to a therapeutic level of fluoride.

TABLE 3 shows the variation in anticaries efficacy with respect tosoluble fluoride content as determined by the ADA method. Using thisexample and the meta-analysis, a 650 ppm fluoride as sodium fluoride wasestimated to effect a reduction in caries of about 29%, or about 25%, orabout 30%, with respect to the placebo or water control in rat cariesexperiments.

TABLE 4 shows the variation in anticaries efficacy with respect toSn-Free antibacterial activity as measured by the Sn-Free iPGRM usingthe dose responsive reduction in rat caries from a well-knownantibacterial, chlorhexidine. These compositions were then analyzedusing iPGRM to define the variation in rat caries effectiveness relativeto its antibacterial efficacy. Since the iPGRM measures reduction inantibacterial efficacy through changes in the pH, non-antibacterialagents that buffer pH, such as sodium bicarbonate, can be corrected forwhen determining the efficacy of the antibacterial agent. This can bedone using placebo controls as appropriate to isolate the Sn-Freeantibacterial contribution. The contribution of Sn was measured throughthe solubility reduction and is excluded from this test. The iPGRM doseresponse of chlorhexidine is shown in FIG. 2.

TABLE 5 shows the variation in anticaries efficacy with respect tocalcium co-ion effect as measured by the iPUM. This method measures thetotal calcium uptake of both soluble and insoluble sources. If aninsoluble source was used, calcium sources were preferred that weresubstantially more soluble than hydroxyapatite so that they willdissolve preferentially when exposed to plaque acids. For example,dicalcium phosphate dihydrate or calcium carbonate dissolve readily onexposure to acid while calcium pyrophosphate is a poor source of calciumfor the purposes here. Insoluble sources have the advantage of increasedresidence time in the plaque and can, therefore, provide a bloom ofcalcium to correspond temporally to the generation of plaque acid. TheiPUM dose response of calcium-containing compositions is shown in FIG.3.

TABLE 6 shows the variation in anticaries efficacy with respect to theF-Free reduction in HAP solubility as measured by F-free HAP. Thecontribution of the fluoride co-ion effect is measured through the ADAmethod and is excluded from this test. An example of the F-free HAPresponse for different levels of Sn in Sn/silica toothpastes is given inFIG. 4.

Finally, the threshold can be defined for which combinations ofsubtherapeutic compositions result in a therapeutic composition. Onaverage, therapeutic compositions can be defined as those providing areduction in caries at least about equivalent to that of a 650 ppmavailable F as NaF/silica toothpaste and/or at least about 24%, 25%,26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, or 34% reduction in caries.Caries reduction and/or anticavity activity can be substantiated usingan actual rat caries performance test. This threshold can be a guideduring development of new compositions for when reductions achieve anapproximately similar therapeutic level of benefit to that of a 650 ppmavailable F as NaF/silica toothpaste with respect to the placebo orwater control.

With these methods, the efficacy can be quantified of variouscompositions along the important anticavity intervention methods. Thesame analysis can be retrospectively applied to a long history of ratcaries experiments conducted during the original development offluoride-containing anticavity compositions. Approximately 170 ratcaries experiments were analyzed containing over 800 individualtreatments for the reductions of caries in two different, but similar,rat caries models. These experiments were conducted between 1959 and2019. Rat caries is the preferred animal model for human caries and isincluded in the US Anticaries Monograph (21 CFR part 355) to ensure theefficacy of fluoride-containing products. It is additionally sensitiveto non-fluoride anticaries mechanisms and compositions. Finally, animalmodels for caries are generally considered interchangeable as long asthey have been properly developed.

This retrospective analysis can help define the scope of the presentinvention. As described herein, the present invention is directed to acombination of subtherapeutic compositions that can result in atherapeutic composition when tested in total. The examples can allow forthe setting of performance thresholds for the subtherapeutic totherapeutic transition in each mechanism as defined by the specifiedperformance test. However, those results, alone, might not anticipatethe combined performance of the compositions in a rat caries experiment.The analysis gave a model from which can predict reductions in ratcaries for combinations of subtherapeutic compositions as describedherein. In combination, the examples, performance thresholds, andmathematical model can help define the present invention.

The retrospective analysis of rat caries experiments was possiblebecause of detailed descriptions of the treatments/ingredients withadditional measurements of F content, Sn content, and pH frequentlydocumented in the experimental record. Efficacy measures in the variousanticaries mechanism methods described above were either measureddirectly when the ingredients could be obtained or estimated using thedetailed description of the treatments and comparisons to similarpresent-day compositions. Estimations of rat caries efficacy using thefour methods named above resulted in a model correlation coefficient,r², of ˜0.76. The correlation coefficient suggests that 76% of thevariation in rat caries efficacy is captured by these methods. Theremaining 24% of variation is typically ascribed to variation commonlyobserved in biological methods. Suffice it to say, we believe thisrepresents a good model for the performance of various compositions inrat caries. The prediction formula is given below for the % reduction inrat caries with respect to water or a silica-based-abrasive,placebo-toothpaste negative control.

% Reduction=1.77*sqrt(ADA/2)+0.146*iPGRM+3.97*iPUM-Ca+0.689*HAP−6.84  Formula 1

The predicted versus actual values for the retrospective analysis of ratcaries data are given in FIG. 5a with a plot of the residuals in FIG. 5b. When a sodium monofluorophosphate (MFP) toothpaste is used, 16 issubtracted from the % Reduction calculated using Formula. This is donebecause MFP is less effective in the rat caries model.

This approach is novel and not obvious to those in the art for manyreasons:

Firstly, there is a long-standing need for a fluoride-free anticavitytoothpaste as evidenced by the distinct lack of such a product in themarket. There are no anti-cavity products in the United States legallymarketed without a therapeutic level of fluoride. This means that no onehas successfully completed a new drug application that meets therigorous FDA standards of safety and efficacy. And as such, there hasbeen a long-felt need for such an alternative.

Secondly, the application methods and model described herein are notenough alone to make a therapeutic composition. Some combinations ofsubtherapeutic compositions are not straightforwardly compatible withone another. For example, soluble fluoride sources are frequentlydeactivated by calcium unless special care is taken. Cationicantibacterial agents can be deactivated by anionic surfactants. The useof stannous ion sources can necessitate the careful choice ofstabilizers and packaging to prevent loss and/or oxidation. Calciumpyrophosphate does not readily dissolve on exposure to plaque acid andis a poor source of calcium for the purposes of protecting teeth fromplaque acids through a co-ion effect. There are many more examples likethis that require expertise in the art to overcome.

Thirdly, nearly all previous examples of anti-cavity compositions in theart focused on how much of a single composition was required to providea therapeutic benefit. None of the examples in the art teach how littleyou can use to achieve a therapeutic benefit from combinations ofsubtherapeutic compositions. This is important in cases where exposureof some ingredients should be limited because of toxicity, aesthetic, orcost concerns. One example comes close by showing a combination of asolubility reducing composition and an antibacterial composition (US2018/0028417 A1); however, it did not teach about the combination ofsubtherapeutic compositions to achieve a therapeutic effect.Additionally, the solubility reducing composition was included at analready therapeutic level to which a second composition enhanced itseffect.

Fourthly, no single example taught a model, method, strategy, orapproach to achieve therapeutic levels of activity through combinationsof subtherapeutic compositions. Furthermore, in none of these artexamples, alone or together, are a method, model, or strategy to achievetherapeutic levels of activity through combinations of subtherapeuticcompositions taught. In some of the disclosed patents, commonsubtherapeutic agents are disclosed; however, never to a degree suchthat a systematic analysis can reveal their impact and interactionleading to a rational design of subtherapeutic compositions. Theperformance of subtherapeutic compositions, excipients, and combinationsthereof are frequently diminished, controlled for experimentally, orjust plainly left uninvestigated. In no case are they explicitlyoptimized for and to the best of our knowledge this has not been done ina systematic way. It is exceedingly unlikely, considering the limitedhistory of rat caries studies published in the art, such an analysiscould be performed by a person of skill in the art by merely evaluatingexisting studies in the published art.

The possible anticaries agents are further described herein.

Definitions

To define more clearly the terms used herein, the following definitionsare provided. Unless otherwise indicated, the following definitions areapplicable to this disclosure. If a term is used in this disclosure butis not specifically defined herein, the definition from the IUPACCompendium of Chemical Terminology, 2nd Ed (1997), can be applied, aslong as that definition does not conflict with any other disclosure ordefinition applied herein, or render indefinite or non-enabled any claimto which that definition is applied.

The term “oral care composition”, as used herein, includes a product,which in the ordinary course of usage, is not intentionally swallowedfor purposes of systemic administration of particular therapeuticagents, but is rather retained in the oral cavity for a time sufficientto contact dental surfaces or oral tissues. Examples of oral carecompositions include dentifrice, tooth gel, subgingival gel, mouthrinse, mousse, foam, mouth spray, lozenge, chewable tablet, chewing gum,tooth whitening strips, floss and floss coatings, breath fresheningdissolvable strips, or denture care or adhesive product. The oral carecomposition may also be incorporated onto strips or films for directapplication or attachment to oral surfaces.

The term “dentifrice composition”, as used herein, includes tooth orsubgingival -paste, gel, or liquid formulations unless otherwisespecified. The dentifrice composition may be a single-phase compositionor may be a combination of two or more separate dentifrice compositions.The dentifrice composition may be in any desired form, such as deepstriped, surface striped, multilayered, having a gel surrounding apaste, or any combination thereof. Each dentifrice composition in adentifrice comprising two or more separate dentifrice compositions maybe contained in a physically separated compartment of a dispenser anddispensed side-by-side.

“Active and other ingredients” useful herein may be categorized ordescribed herein by their cosmetic and/or therapeutic benefit or theirpostulated mode of action or function. However, it is to be understoodthat the active and other ingredients useful herein can, in someinstances, provide more than one cosmetic and/or therapeutic benefit orfunction or operate via more than one mode of action. Therefore,classifications herein are made for the sake of convenience and are notintended to limit an ingredient to the particularly stated function(s)or activities listed.

The term “orally acceptable carrier” comprises one or more compatiblesolid or liquid excipients or diluents which are suitable for topicaloral administration. By “compatible,” as used herein, is meant that thecomponents of the composition are capable of being commingled withoutinteraction in a manner which would substantially reduce thecomposition's stability and/or efficacy. The carriers or excipients ofthe present invention can include the usual and conventional componentsof mouthwashes or mouth rinses, as more fully described hereinafter:Mouthwash or mouth rinse carrier materials typically include, but arenot limited to one or more of water, alcohol, humectants, surfactants,and acceptance improving agents, such as flavoring, sweetening, coloringand/or cooling agents.

The term “substantially free” as used herein refers to the presence ofno more than 0.05%, preferably no more than 0.01%, and more preferablyno more than 0.001%, of an indicated material in a composition, by totalweight of such composition.

The term “essentially free” as used herein means that the indicatedmaterial is not deliberately added to the composition, or preferably notpresent at analytically detectable levels. It is meant to includecompositions whereby the indicated material is present only as animpurity of one of the other materials deliberately added.

The term “therapeutic anticaries activity”, as used herein, is theanticaries activity provided by a composition including an anticariesdrug and/or one or more anticaries agents in a therapeutic dose. Atherapeutic dose for fluoride ions is defined in the United States bythe Food and Drug Administration (FDA) Monograph (21 CFR part 355). Forexample, a therapeutic amount of sodium fluoride in a paste dosage form(i.e. paste dentifrice) is 850 to 1,150 ppm with an available fluorideion concentration of at least 650 ppm. Other anticaries drugs aredisclosed in 21 CFR part 355, which is herein incorporated by reference.Other therapeutic dosages are available in respective jurisdictions.Thus, as used herein, therapeutic anticaries activity for a compositioncomprising one or more anticaries agents is the anticaries benefit whichis equivalent or better than the anticaries benefit provided by acomposition comprising sodium fluoride with at least 650 ppm ofavailable fluoride ions.

The term “subtherapeutic anticaries activity”, as used herein, is theanticaries activity of a composition that has a lower anticaries benefitrelative to the anticaries benefit provided by a composition comprisingsodium fluoride with at least 650 ppm of available fluoride ions, asdefined in “therapeutic anticaries drug composition.” The term“subtherapeutic anticaries activity” also can also describe anticariesagents that can, in combination with additional anticaries agents, leadto a therapeutic benefit relative to the anticaries benefit provided bya composition comprising sodium fluoride with at least 650 ppm ofavailable fluoride ions, when utilized in an oral care compositionsuitable for use in the oral cavity of a human, but alone has not beenshown to provide a therapeutic benefit at concentrations suitable foruse in an oral care composition.

The term “anticaries drug”, as used herein, is a drug that aids in theprevention and prophylactic treatment of dental cavities (decay,caries). This can include fluoride ion sources, such as in 21 CFR part355 and anticaries agents, as described herein.

While compositions and methods are described herein in terms of“comprising” various components or steps, the compositions and methodscan also “consist essentially of” or “consist of” the various componentsor steps, unless stated otherwise.

As used herein, the word “or” when used as a connector of two or moreelements is meant to include the elements individually and incombination; for example, X or Y, means X or Y or both.

As used herein, the articles “a” and “an” are understood to mean one ormore of the material that is claimed or described, for example, “an oralcare composition” or “a bleaching agent.”

All measurements referred to herein are made at about 23° C. (i.e. roomtemperature) unless otherwise specified.

Generally, groups of elements are indicated using the numbering schemeindicated in the version of the periodic table of elements published inChemical and Engineering News, 63(5), 27, 1985. In some instances, agroup of elements can be indicated using a common name assigned to thegroup; for example, alkali metals for Group 1 elements, alkaline earthmetals for Group 2 elements, and so forth.

Several types of ranges are disclosed in the present invention. When arange of any type is disclosed or claimed, the intent is to disclose orclaim individually each possible number that such a range couldreasonably encompass, including end points of the range as well as anysub-ranges and combinations of sub-ranges encompassed therein.

The term “about” means that amounts, sizes, formulations, parameters,and other quantities and characteristics are not and need not be exact,but can be approximate and/or larger or smaller, as desired, reflectingtolerances, conversion factors, rounding off, measurement errors, andthe like, and other factors known to those of skill in the art. Ingeneral, an amount, size, formulation, parameter or other quantity orcharacteristic is “about” or “approximate” whether or not expresslystated to be such. The term “about” also encompasses amounts that differdue to different equilibrium conditions for a composition resulting froma particular initial mixture. Whether or not modified by the term“about,” the claims include equivalents to the quantities. The term“about” can mean within 10% of the reported numerical value, preferablywithin 5% of the reported numerical value.

The dentifrice composition can be in any suitable form, such as a solid,liquid, powder, paste, or combinations thereof. The oral carecomposition can be dentifrice, tooth gel, subgingival gel, mouth rinse,mousse, foam, mouth spray, lozenge, chewable tablet, chewing gum, toothwhitening strips, floss and floss coatings, breath fresheningdissolvable strips, or denture care or adhesive product. The componentsof the dentifrice composition can be incorporated into a film, a strip,a foam, or a fiber-based dentifrice composition.

Section headers are provided below for organization and convenienceonly. The section headers do not suggest that a compound cannot bewithin more than one section. In fact, compounds can fall within morethan one section. For example, stannous chloride can be both a tin ionsource and a surface adsorbed stabilizer, stannous fluoride can be botha tin ion source and a fluoride ion source, glycine can be an aminoacid, a buffering agent, and/or a surface adsorbed stabilizer, amongnumerous other compounds that can fit amongst several categories and/orsections.

Anticaries Activity

The oral care compositions, as described herein, can comprise one ormore anticaries agents, which collectively demonstrate therapeuticanticaries activity. As described herein, therapeutic anticariesactivity is defined by the relevant regulatory agency in a jurisdictionof interest, such as the U.S. FDA in the United States. At the FDA, atherapeutic amount of sodium fluoride in a paste dosage form (i.e. pastedentifrice) is 850 to 1,150 ppm with an available fluoride ionconcentration of at least 650 ppm. Thus, the oral care compositions ofthe present invention can have an anticaries benefit at least about ofthe anticaries benefit of a composition comprising at least, at leastabout, or greater than 650 ppm, 800 ppm, 850 ppm, 1100 ppm, 1150 ppm,1450 ppm, and/or 2800 ppm of free fluoride ions, which can correspond totherapeutic anticaries activity.

The anticaries activity of the oral care compositions, as describedherein, can also be described by a rat caries score. The oral carecompositions can have a rat caries score of about 35 or less, about 30or less, about 25 or less, and/or about 20 or less. The anticariesactivity of the oral care compositions, as described herein, can also bedescribed the percent reduction of the rat caries score relative to aplacebo toothpaste. The oral care compositions can have a percentreduction in caries relative to a placebo toothpaste (i.e. a negativecontrol) of at least about 25%, at least about 29%, at least about 30%,at least about 35%, and/or at least about 40%.

The anticaries activity of the oral care compositions, as describedherein, can also be described by a percent reduction of the rat cariesscore relative to an oral care composition comprising a therapeutic doseof an anticaries agent (i.e. a positive control, such as USP NaF). Theoral care compositions can have a percent reduction in caries relativeto a positive control, such as USP NaF, of about 50% or more, about 60%or more, about 65% or more, about 70% or more, about 75% or more, about80% or more, about 90% or more, about 100% or more, about 125% or more,and/or about 150% or more.

Anticaries Agent

The oral care composition, as described herein, comprise one or moreanticaries compositions, the one or more anticaries compositionscomprising one or more anticaries agents, which can, individually, be ina therapeutic dose or a subtherapeutic dose.

The oral care composition can comprise a first subtherapeutic anticariesagent and a second subtherapeutic anticaries agent, which in total leadto a therapeutic anticaries benefit. The first and second subtherapeuticanti caries agents can be fluoride-free, as described herein, or one ofthe subtherapeutic agents can comprise a subtherapeutic amount offluoride ions.

The oral care composition can comprise a therapeutic anticaries agentcomprising a fluoride ion source and a subtherapeutic anticaries agent,which is free from a fluoride ion source, where the overall oral carecomposition has a higher anticaries benefit than the anticaries benefitassociated with the a therapeutic anticaries composition alone. Theanticaries activity of a composition comprising a therapeutic anticariesagent and a subtherapeutic anticaries agent can allow for anticariesactivity that is normally available with a prescription strengthconcentration of fluoride ions.

The anticaries agent can be active against caries through one of thesefour mechanisms: i) suppressing acid formation via antibacterial action;ii) reducing enamel solubility through a calcium co-ion effect; iii)reducing enamel solubility through a fluoride co-ion effect; and iv)reducing enamel solubility through surface adsorbed stabilizers. Thus,the anticaries agent can be an antibacterial agent, a calcium ionsource, a fluoride ion source, a surface adsorbed stabilizer. However, acompound can fall within more than one of these categories, such as, forexample, stannous chloride, which can be an antibacterial agent and asurface adsorbed stabilizer or stannous fluoride, which can be anantibacterial agent, a fluoride ion source, and a surface adsorbedstabilizer.

Antibacterial Agent

The oral care composition can comprise one or more anticaries agents,the one or more anticaries agents can comprise one or more antibacterialagents. The antibacterial agent can be any agent that suppresses acidformation by the bacteria of dental caries. Suitable antibacterialagents include agents that those that can provide at least about an 80%,or about 30%, 60%, 65%, 75%, 85%, 90%, or 95%, reduction in ΔpH withrespect to Crest® Cavity Protection that thereby reduce caries at leastabout 9%, or about 1%, 6%, 7%, 8%, 10%, 11%, or 12%, with respect to theplacebo or water control in rat caries experiments.

Suitable antibacterial agents include hops acids, such as hops alphaacids, hops beta acids, hydrogenated hops acids, and/or combinationsthereof. Other suitable antibacterial agents include metal ion sources,such as tin ion sources, zinc ion sources, copper ion sources, and/orcombinations thereof. Other suitable antibacterial agents includetriclosan, extracts from any species within the genus Magnolia, extractsfrom any species within the genus Humulus. Other suitable antibacterialagents include hops acids, tin ion sources, benzyl alcohol, sodiumbenzoate, menthylglycyl acetate, menthyl lactate, L-menthol,o-neomenthol, chlorophyllin copper complex, phenol, oxyquinoline, and/orcombinations thereof. Other suitable antibacterial agents include one ormore amino acids, such as basic amino acids.

The oral care composition can comprise from about 0.01% to about 10%,from about 1% to about 5%, or from about 0.5% to about 15% of anantibacterial agent. Some, but not all, suitable antibacterial agentswill be discussed separately.

Surface Adsorbed Stabilizers

The oral care composition can comprise one or more anticaries agents,the one or more anticaries agents can comprise one or more surfaceadsorbed stabilizers. The surface adsorbed stabilizer can be any agentthat can adsorb on an enamel surface. A suitable surface adsorbedstabilizer can be any compound that can provide at least a 17%, at leasta 5%, or at least a 20%, at least a 30%, and/or at least a 15%,reduction in solubility relative to water in the F-free HAP dissolutionmethod, which thereby reduces caries at least 5%, 6%, 7%, 8%, 9%, 10%,11%, 12%, 13%, and/or 18% with respect to the placebo or water controlin rat caries experiments.

Suitable surface adsorbed stabilizers include metal ion sources, such astin ion sources, zinc ion sources, copper ion sources, aluminum ionsources, titanium ion sources and/or combinations thereof. Othersuitable surface adsorbed stabilizers include bioactive materials, aminoacids, and/or combinations thereof. Some, but not all, suitable surfaceadsorbed stabilizes will be discussed separately.

Humulus lupulus

The oral care compositions of the present invention can comprise atleast one hops compound from Formula I and/or Formula IV. The compoundfrom Formula I and/or Formula IV can be provided by any suitable source,such as an extract from Humulus lupulus or Hops, Humulus lupulus itself,a synthetically derived compound, and/or salts, prodrugs, or otheranalogs thereof. The hops extract can comprise one or more hops alphaacids, one or more hops iso-alpha acids, one or more hops beta acids,one or more hops oils, one or more flavonoids, one or more solvents,and/or water. Suitable hops alpha acids (generically shown in Formula I)can include humulone (Formula II), adhumulone, cohumulone, posthumulone,prehumulone, and/or mixtures thereof. Suitable hops iso-alpha acids caninclude cis-isohumulone and/or trans-isohumulone. The isomerization ofhumulone into cis-isohumulone and trans-isohumulone can be representedby Formula III.

A is the acidic hydroxyl functional group in the alpha position, B arethe acidic hydroxyl functional groups in the beta position, and R is analkyl functional group.

Suitable hops beta acids can include lupulone, adlupulone, colupulone,and/or mixtures thereof. A suitable hops beta acid can include acompound a described in Formula IV, V, VI, and/or VII.

B are the acidic hydroxyl functional groups in the beta position and Ris an alkyl functional group.

While hops alpha acids can demonstrate some antibacterial activity, hopsalpha acids also have a bitter taste. The bitterness provided by hopsalpha acids can be suitable for beer, but are not suitable for use inoral care compositions. In contrast, hops beta acids can be associatedwith a higher antibacterial and/or anticaries activity, but not asbitter a taste. Thus, a hops extract with a higher proportion of betaacids to alpha acids than normally found in nature, can be suitable foruse in oral care compositions for use as an antibacterial and/oranticaries agent.

A natural hops source can comprise from about 2% to about 12%, by weightof the hops source, of hops beta acids depending on the variety of hops.Hops extracts used in other contexts, such as in the brewing of beer,can comprise from about 15% to about 35%, by weight of the extract, ofhops beta acids. The hops extract desired herein can comprise at leastabout 35%, at least about 40%, at least about 45%, from about 35% toabout 95%, from about 40% to about 90%, or from about 45% to about 99%,of hops beta acids. The hops beta acids can be in an acidic form (i.e.with attached hydrogen atom(s) to the hydroxl functional group(s)) or asa salt form.

A suitable hops extract is described in detail in U.S. Pat. No.7,910,140, which is herein incorporated by reference in its entirety.The hops beta acids desired can be non-hydrogenated, partiallyhydrogenated by a non-naturally occurring chemical reaction, orhydrogenated by a non-naturally occurring chemical reaction. The hopsbeta acid can be essentially free of or substantially free ofhydrogenated hops beta acid and/or hops acid. A non-naturally occurringchemical reaction is a chemical reaction that was conducted with the aidof chemical compound not found within Humulus lupulus, such as achemical hydrogenation reaction conducted with high heat not normallyexperienced by Humulus lupulus in the wild and/or a metal catalyst.

A natural hops source can comprise from about 2% to about 12%, by weightof the hops source, of hops alpha acids. Hops extracts used in othercontexts, such as in the brewing of beer, can comprise from about 15% toabout 35%, by weight of the extract, of hops alpha acids. The hopsextract desired herein can comprise less than about 10%, less than about5%, less than about 1%, or less than about 0.5%, by weight of theextract, of hops alpha acids.

Hops oils can include terpene hydrocarbons, such as myrcene, humulene,caryophyllene, and/or mixtures thereof. The hops extract desired hereincan comprise less than 5%, less than 2.5%, or less than 2%, by weight ofthe extract, of one or more hops oils.

Flavonoids present in the hops extract can include xanthohumol,8-prenylnaringenin, isoxanthohumol, and/or mixtures thereof. The hopsextract can be substantially free of, essentially free of, free of, orhave less than 250 ppm, less than 150 ppm, and/or less than 100 ppm ofone or more flavonoids.

As described in U.S. Pat. No. 5,370,863, hops acids have been previouslyadded to oral care compositions. However, the oral care compositionstaught by U.S. Pat. No. 5,370,863 only included up to 0.01%, by weightof the oral care composition. While not wishing to be bound by theory,it is believed that U.S. Pat. No. 5,370,863 could only incorporate a lowamount of hops acids because of the bitterness of hops alpha acids. Ahops extract with a low level of hops alpha acids would not have thisconcern.

The hops compound can be combined with or free from an extract fromanother plant, such as a species from genus Magnolia. The hops compoundscan be combined with or free from triclosan.

The oral care composition can comprise from about 0.01% to about 10%,greater than 0.01% to about 10%, from about 0.05%, to about 10%, fromabout 0.1% to about 10%, from about 0.2% to about 10%, from about 0.2%to about 10%, from about 0.2% to about 5%, from about 0.25% to about 2%,from about 0.05% to about 2%, or from greater than 0.25% to about 2%, ofhops beta acid, as described herein. The hops beta acids can be providedby a suitable hops extract, the hops plant itself, or a syntheticallyderived compound. The hops beta acid can be provided as neutral, acidiccompounds, and/or as salts with a suitable counter ion, such as sodium,potassium, ammonia, or any other suitable counter ion.

The hops beta acid can be provided by a hops extract, such as an extractfrom Humulus lupulus with at least 35%, by weight of the extract, ofhops beta acid and less than 1%, by weight of the hops extract, of hopsalpha acid. The oral care composition can comprise 0.01% to about 10%,greater than 0.01% to about 10%, from about 0.05%, to about 10%, fromabout 0.1% to about 10%, from about 0.2% to about 10%, from about 0.2%to about 10%, from about 0.2% to about 5%, from about 0.25% to about 2%,from about 0.05% to about 2%, or from greater than 0.25% to about 2%, ofhops extract, as described herein.

Fluoride Ion Source

The oral care composition can comprise fluoride. Fluoride can beprovided by a fluoride ion source. The fluoride ion source can compriseone or more fluoride containing compounds, such as stannous fluoride,sodium fluoride, titanium fluoride, calcium fluoride, calcium phosphatesilicate fluoride, potassium fluoride, amine fluoride, sodiummonofluorophosphate, zinc fluoride, and/or mixtures thereof.

The fluoride ion source and the tin ion source can be the same compound,such as for example, stannous fluoride, which can generate tin ions andfluoride ions. Additionally, the fluoride ion source and the tin ionsource can be separate compounds, such as when the tin ion source isstannous chloride and the fluoride ion source is sodiummonofluorophosphate or sodium fluoride.

The fluoride ion source and the zinc ion source can be the samecompound, such as for example, zinc fluoride, which can generate zincions and fluoride ions. Additionally, the fluoride ion source and thezinc ion source can be separate compounds, such as when the zinc ionsource is zinc phosphate and the fluoride ion source is stannousfluoride.

The fluoride ion source can be essentially free of, substantially freeof, or free of stannous fluoride. Thus, the oral care composition cancomprise sodium fluoride, potassium fluoride, amine fluoride, sodiummonofluorophosphate, zinc fluoride, and/or mixtures thereof.

The oral care composition can comprise a fluoride ion source capable ofproviding from about 50 ppm to about 5000 ppm, and preferably from about500 ppm to about 3000 ppm of free fluoride ions. To deliver the desiredamount of fluoride ions, the fluoride ion source may be present in theoral care composition at an amount of from about 0.0025% to about 5%,from about 0.01% to about 10%, from about 0.2% to about 1%, from about0.5% to about 1.5%, or from about 0.3% to about 0.6%, by weight of theoral care composition. Alternatively, the oral care composition cancomprise less than 0.1%, less than 0.01%, be essentially free of, besubstantially free of, or free of a fluoride ion source.

A subtherapeutic amount of a fluoride ion source for the purposes ofanticaries activity can include an oral care composition comprising lessthan 1100 ppm, less than 800 ppm, less than 650 ppm, less than 500 ppm,equal to and less than 500 ppm, less than 250 ppm, equal to and lessthan 250 ppm, and/or combinations thereof. A subtherapeutic amount of afluoride ion source has a measurable anticaries affect, such as at leasta 5%, at least a 7.5%, at least a 10%, at least a 15%, at least a 20%,and/or combinations thereof, but not a therapeutic anticaries effect asrequired by a regulatory agency in any jurisdiction of interest, asdescribed further herein.

Tin Ion Source

The oral care composition of the present invention can comprise tin,such as from a tin ion source. The tin ion source can be any suitablecompound that can provide tin ions in an oral care composition and/ordeliver tin ions to the oral cavity when the dentifrice composition isapplied to the oral cavity. The tin ion source can comprise one or moretin containing compounds, such as stannous fluoride, stannous chloride,stannous bromide, stannous iodide, stannous oxide, stannous oxalate,stannous sulfate, stannous sulfide, stannic fluoride, stannic chloride,stannic bromide, stannic iodide, stannic sulfide, and/or mixturesthereof. Tin ion source can comprise stannous fluoride, stannouschloride, and/or mixture thereof. The tin ion source can also be afluoride-free tin ion source, such as stannous chloride.

The oral care composition can comprise from about 0.0025% to about 5%,from about 0.01% to about 10%, from about 0.2% to about 1%, from about0.5% to about 1.5%, or from about 0.3% to about 0.6%, by weight of theoral care composition, of a tin ion source.

Ca Ion Source

The oral care composition of the present invention can comprise calcium,such as from a calcium ion source. The calcium ion source can be anysuitable compound or molecule that can provide calcium ions in an oralcare composition and/or deliver calcium ions to the oral cavity when theoral care composition is applied to the oral cavity. The calcium ionsource can comprise a calcium salt, a calcium abrasive, and/orcombinations thereof. In some cases, a calcium salt may also beconsidered a calcium abrasive or a calcium abrasive may also beconsidered a calcium salt.

The calcium ion source can comprise a calcium abrasive. The calciumabrasive can be any suitable abrasive compound that can provide calciumions in an oral care composition and/or deliver calcium ions to the oralcavity when the oral care composition is applied to the oral cavity. Thecalcium abrasive can comprise one or more calcium abrasive compounds,such as calcium carbonate, precipitated calcium carbonate (PCC), groundcalcium carbonate (GCC), chalk, dicalcium phosphate, calciumpyrophosphate, and/or mixtures thereof.

The calcium ion source can comprise a calcium salt, or a compound thatcan provide calcium ions in an oral care composition and/or delivercalcium ions to the oral cavity when the oral care composition isapplied to the oral cavity that can not act as an abrasive. The calciumsalt can comprise one or more calcium compounds, such as calciumchloride, calcium nitrate, calcium phosphate, calcium lactate, calciumoxalate, calcium oxide, calcium gluconate, calcium citrate, calciumbromide, calcium iodate, calcium iodide, hydroxyapatite, fluorapatite,calcium sulfate, calcium glycerophosphate, and/or combinations thereof.

The calcium ion source can have a iPUM-Ca of at least 2 times, at least4 times, at least 6 times of the iPUM-Ca of untreated biofilm asdescribed herein.

The oral care composition can comprise from about 5% to about 70%, fromabout 10% to about 50%, from about 10% to about 60%, from about 20% toabout 50%, from about 25% to about 40%, or from about 1% to about 50% ofa calcium ion source.

Buffering Agent

The oral care composition can comprise a buffering agent. The bufferingagent can be a weak acid or base that can maintain a particular pH at aselected site in the oral cavity. For example, the buffering agent canmaintain a pH at a tooth's surface to mitigate the impact of plaqueacids produced by bacteria. The buffering agent can comprise a conjugateacid of an ion also present in the oral care composition. For example,if the calcium ion source comprises calcium carbonate, the bufferingagent can comprise a bicarbonate anion (—HCO₃ ⁻). The buffering agentcan comprise a conjugate acid/base pair, such as citric acid and sodiumcitrate.

Suitable buffering systems can include phosphate, citrate salts,carbonate/bicarbonate salts, a tris buffer, imidazole, urea, borate,and/or combinations thereof. Suitable buffering agents includebicarbonate salts, such as sodium bicarbonate, glycine, orthophosphate,arginine, urea, and or/combinations thereof.

The oral care composition can comprise from about 0.5% to about 30%,from about 5% to about 25% or from about 10% to about 20%, of one ormore buffering agents.

Biofilm Modifier

The oral care composition can comprise one or more biofilm modifiers. Abiofilm modifier can comprise a polyol, an ammonia generating compound,and/or a glucosyltransferase inhibitor.

A polyol is an organic compound with more than one hydroxyl functionalgroups. The polyol can be any suitable compound that can weaklyassociate, interact, or bond to tin ions while the oral care compositionis stored prior to use. The polyol can be a sugar alcohol, which areaclass of polyols that can be obtained through the hydrogenation of sugarcompounds with the formula (CHOH)_(n)H₂. The polyol can be glycerin,erythritol, xylitol, sorbitol, mannitol, butylene glycol, lactitol,and/or combinations thereof. The oral care composition can comprise0.01% to about 70%, from about 5% to about 70%, from about 5% to about50%, from about 10% to about 60%, from about 10% to about 25%, or fromabout 20% to about 80%, by weight of the oral care composition, of apolyol.

The ammonia generating compound can be any suitable compound that cangenerate ammonia upon delivery to the oral cavity. Suitable ammoniagenerating compounds include arginine, urea, and/or combinationsthereof. The oral care composition can comprise from about 0.01% toabout 10%, from about 1% to about 5%, or from about 1% to about 25% ofone or more ammonia generating compounds.

The glucosyltransferase inhibitor can be any suitable compound that caninhibit a glucosyltransferase. Glucosyltransferases are enzymes that canestablish natural glycosidic linkages. In particular, these enzymesbreak down poly- or oligosaccharide moieties into simple sugars forbacteria associated with dental caries. As such, any compound that caninhibit this process can help prevent dental caries. Suitableglucosyltransferase inhibitors include oleic acid, epicatechin, tannins,tannic acid, moenomycin, caspofungin, ethambutol, lufenuron, and/orcombinations thereof. The oral care composition can comprise from about0.001% to about 5%, from about 0.01% to about 2%, or about 1% of one ormore glucosyltransferase inhibitors.

Metal Ion Source

The oral care composition can comprise metal, such as from a metal ionsource comprising one or more metal ions. The metal ion source cancomprise or be in addition to the tin ion source and/or the zinc ionsource, as described herein. Suitable metal ion sources includecompounds with metal ions, such as, but not limited to Sn, Zn, Cu, Mn,Mg, Sr, Ti, Fe, Mo, B, Ba, Ce, Al, In and/or mixtures thereof. The tracemetal source can be any compound with a suitable metal and anyaccompanying ligands and/or anions.

Suitable ligands and/or anions that can be paired with metal ion sourcesinclude, but are not limited to acetate, ammonium sulfate, benzoate,bromide, borate, carbonate, chloride, citrate, gluconate,glycerophosphate, hydroxide, iodide, oxide, propionate, D-lactate,DL-lactate, orthophosphate, pyrophosphate, sulfate, nitrate, tartrate,and/or mixtures thereof.

The oral care composition can comprise from about 0.01% to about 10%,from about 1% to about 5%, or from about 0.5% to about 15% of a metalion source.

Bioactive Materials

The oral care composition can also include bioactive materials suitablefor the remineralization of a tooth. Suitable bioactive materialsinclude bioactive glasses, Novamin™, Recaldent™, hydroxyapatite, one ormore amino acids, such as, for example, arginine, citrulline, glycine,lysine, or histidine, or combinations thereof. Suitable examples ofcompositions comprising arginine are found in U.S. Pat. Nos. 4,154,813and 5,762,911, which are herein incorporated by reference in theirentirety. Other suitable bioactive materials include any calciumphosphate compound. Other suitable bioactive materials include compoundscomprising a calcium source and a phosphate source.

Amino acids are organic compounds that contain an amine functionalgroup, a carboxyl functional group, and a side chain specific to eachamino acid. Suitable amino acids include, for example, amino acids witha positive or negative side chain, amino acids with an acidic or basicside chain, amino acids with polar uncharged side chains, amino acidswith hydrophobic side chains, and/or combinations thereof. Suitableamino acids also include, for example, arginine, histidine, lysine,aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine,cysteine, selenocysteine, glycine, proline, alanine, valine, isoleucine,leucine, methionine, phenylalanine, tyrosine, tryptophan, citrulline,ornithine, creatine, diaminobutonic acid, diaminoproprionic acid, saltsthereof, and/or combinations thereof.

Bioactive glasses are comprising calcium and/or phosphate which can bepresent in a proportion that is similar to hydroxyapatite. These glassescan bond to the tissue and are biocompatible. Bioactive glasses caninclude a phosphopeptide, a calcium source, phosphate source, a silicasource, a sodium source, and/or combinations thereof.

The oral care composition can comprise from about 0.01% to about 20%,from about 0.1% to about 10%, or from about 1% to about 10% of abioactive material by weight of the oral care composition.

Abrasive

The oral care composition can comprise a calcium abrasive, as describedherein, and/or a non-calcium abrasive, such as bentonite, silica gel (byitself, and of any structure), precipitated silica, amorphousprecipitated silica (by itself, and of any structure as well), hydratedsilica, perlite, titanium dioxide, calcium pyrophosphate, dicalciumphosphate dihydrate, alumina, hydrated alumina, calcined alumina,aluminum silicate, insoluble sodium metaphosphate, insoluble potassiummetaphosphate, insoluble magnesium carbonate, zirconium silicate,particulate thermosetting resins and other suitable abrasive materials.Such materials can be introduced into the oral care compositions totailor the polishing characteristics of the target dentifriceformulation. The oral care composition can comprise from about 5% toabout 70%, from about 10% to about 50%, from about 10% to about 60%,from about 20% to about 50%, from about 25% to about 40%, or from about1% to about 50%, by weight of the oral care composition, of thenon-calcium abrasive.

Alternatively, the oral care composition can be substantially free of,essentially free of, or free of silica, alumina, or any othernon-calcium abrasive. The oral care composition can comprise less thanabout 5%, less than about 1%, less than about 0.5%, less than about0.1%, or 0% of a non-calcium abrasive, such as silica and/or alumina.

Water

The oral care composition of the present invention can be anhydrous, alow water formulation, or a high water formulation. In total, the oralcare composition can comprise from 0% to about 99%, from about 5% toabout 75%, about 20% or greater, about 30% or greater, or about 50% orgreater by weight of the composition, of water. Preferably, the water isUSP water.

In a high water oral care composition and/or toothpaste formulation, theoral care composition comprises from about 45% to about 75%, by weightof the composition, of water. The high water oral care compositionand/or toothpaste formulation can comprise from about 45% to about 65%,from about 45% to about 55%, or from about 46% to about 54%, by weightof the composition, of water. The water may be added to the high waterformulation and/or may come into the composition from the inclusion ofother ingredients.

In a low water oral care composition and/or toothpaste formulation, theoral care composition comprises from about 5% to about 45%, by weight ofthe composition, of water. The low water oral care composition cancomprise from about 5% to about 35%, from about 10% to about 25%, orfrom about 20% to about 25%, by weight of the composition, of water. Thewater may be added to the low water formulation and/or may come into thecomposition from the inclusion of other ingredients.

In an anhydrous oral care composition and/or toothpaste formulation, theoral care composition comprises less than about 10%, by weight of thecomposition, of water. The anhydrous composition comprises less thanabout 5%, less than about 1%, or 0%, by weight of the composition, ofwater. The water may be added to the anhydrous formulation and/or maycome into the composition from the inclusion of other ingredients.

A mouth rinse formulation comprises from about 75% to about 99%, fromabout 75% to about 95%, or from about 80% to about 95% of water.

The composition can also comprise other orally acceptable carriermaterials, such as alcohol, humectants, polymers, surfactants, andacceptance improving agents, such as flavoring, sweetening, coloringand/or cooling agents.

pH

The pH of the disclosed composition can be from about 4 to about 10,from about 7 to about 10, greater than 7 to about 10, greater than 8 toabout 10, greater than 7, greater than 7.5, greater than 8, greater than9, or from about 8.5 to about 10.

Zinc Ion Source

The oral care composition can comprise zinc, such as from a zinc ionsource. The zinc ion source can comprise one or more zinc containingcompounds, such as zinc fluoride, zinc lactate, zinc oxide, zincphosphate, zinc chloride, zinc acetate, zinc hexafluorozirconate, zincsulfate, zinc tartrate, zinc gluconate, zinc citrate, zinc malate, zincglycinate, zinc pyrophosphate, zinc metaphosphate, zinc oxalate, and/orzinc carbonate. The zinc ion source can be a fluoride-free zinc ionsource, such as zinc phosphate, zinc oxide, and/or zinc citrate.

The zinc ion source may be present in the total oral care composition atan amount of from about 0.01% to about 10%, from about 0.2% to about 1%,from about 0.5% to about 1.5%, or from about 0.3% to about 0.6%, byweight of the dentifrice composition.

Polyphosphates

The oral care composition can comprise polyphosphate, such as from apolyphosphate source. A polyphosphate source can comprise one or morepolyphosphate molecules. Polyphosphates are a class of materialsobtained by the dehydration and condensation of orthophosphate to yieldlinear and cyclic polyphosphates of varying chain lengths. Thus,polyphosphate molecules are generally identified with an average number(n) of polyphosphate molecules, as described below. A polyphosphate isgenerally understood to consist of two or more phosphate moleculesarranged primarily in a linear configuration, although some cyclicderivatives may be present.

Preferred polyphosphates are those having an average of two or morephosphate groups so that surface adsorption at effective concentrationsproduces sufficient non-bound phosphate functions, which enhance theanionic surface charge as well as hydrophilic character of the surfaces.Preferred in this invention are the linear polyphosphates having theformula: XO(XPO₃)_(n)X, wherein X is sodium, potassium, ammonium, or anyother alkali metal cations and n averages from about 2 to about 21.Alkali earth metal cations, such as calcium, are not preferred becausethey tend to form insoluble fluoride salts from aqueous solutionscomprising a fluoride ions and alkali earth metal cations. Thus, theoral care compositions disclosed herein can be free of, essentially freeof, or substantially free of calcium pyrophosphate.

Some examples of suitable polyphosphate molecules include, for example,pyrophosphate (n=2), tripolyphosphate (n=3), tetrapolyphosphate (n=4),sodaphos polyphosphate (n=6), hexaphos polyphosphate (n=13), benephospolyphosphate (n=14), hexametaphosphate (n=21), which is also known asGlass H. Polyphosphates can include those polyphosphate compoundsmanufactured by FMC Corporation, ICL Performance Products, and/orAstaris.

The oral care composition can comprise from about 0.01% to about 15%,from about 0.1% to about 10%, from about 0.5% to about 5%, from about 1to about 20%, or about 10% or less, by weight of the oral carecomposition, of the polyphosphate source.

Humectants

The oral care composition can comprise one or more humectants, have lowlevels of a humectant, be substantially free of, substantially free of,or be free of a humectant. Humectants serve to add body or “mouthtexture” to an oral care composition or dentifrice as well as preventingthe dentifrice from drying out. Suitable humectants include polyethyleneglycol (at a variety of different molecular weights), propylene glycol,glycerin (glycerol), erythritol, xylitol, sorbitol, mannitol, butyleneglycol, lactitol, hydrogenated starch hydrolysates, and/or mixturesthereof. The oral care composition can comprise one or more humectantseach at a level of from 0 to about 70%, from about 5% to about 50%, fromabout 10% to about 60%, or from about 20% to about 80%, by weight of theoral care composition.

Surfactants

The oral care composition can comprise one or more surfactants. Thesurfactants can be used to make the compositions more cosmeticallyacceptable. The surfactant is preferably a detersive material whichimparts to the composition detersive and foaming properties. Suitablesurfactants are safe and effective amounts of anionic, cationic,nonionic, zwitterionic, amphoteric and betaine surfactants.

Suitable anionic surfactants include, for example, the water solublesalts of alkyl sulfates having from 8 to 20 carbon atoms in the alkylradical and the water-soluble salts of sulfonated monoglycerides offatty acids having from 8 to 20 carbon atoms. Sodium lauryl sulfate(SLS) and sodium coconut monoglyceride sulfonates are examples ofanionic surfactants of this type. Other suitable anionic surfactantsinclude sarcosinates, such as sodium lauroyl sarcosinate, taurates,sodium lauryl sulfoacetate, sodium lauroyl isethionate, sodium laurethcarboxylate, and sodium dodecyl benzene sulfonate. Combinations ofanionic surfactants can also be employed.

Another suitable class of anionic surfactants are alkyl phosphates. Thesurface active organophosphate agents can have a strong affinity forenamel surface and have sufficient surface binding propensity to desorbpellicle proteins and remain affixed to enamel surfaces. Suitableexamples of organophosphate compounds include mono-, di- or triestersrepresented by the general structure below wherein Z₁, Z₂, or Z₃ may beidentical or different with at least one being an organic moiety. Z₁,Z₂, or Z₃ can be selected from linear or branched, alkyl or alkenylgroup of from 1 to 22 carbon atoms, optionally substituted by one ormore phosphate groups; alkoxylated alkyl or alkenyl, (poly)saccharide,polyol or polyether group.

Some other agents include alkyl or alkenyl phosphate esters representedby the following structure:

wherein R₁ represents a linear or branched, alkyl or alkenyl group offrom 6 to 22 carbon atoms, optionally substituted by one or morephosphate groups; n and m, are individually and separately, 2 to 4, anda and b, individually and separately, are 0 to 20; Z and Z may beidentical or different, each represents hydrogen, alkali metal,ammonium, protonated alkyl amine or protonated functional alkylamine,such as analkanolamine, or a R—(OCH2)(OCH)— group. Examples of suitableagents include alkyl and alkyl (poly)alkoxy phosphates such as laurylphosphate; PPGS ceteareth-10 phosphate; laureth-1 phosphate; laureth-3phosphate; laureth-9 phosphate; trilaureth-4 phosphate; C₁₂₋₁₈ PEG 9phosphate: and sodium dilaureth-10 phosphate. The alkyl phosphate can bepolymeric. Examples of polymeric alkyl phosphates include thosecontaining repeating alkoxy groups as the polymeric portion, inparticular 3 or more ethoxy, propoxy isopropoxy or butoxy groups.

Other suitable anionic surfactants are sarcosinates, isethionates andtaurates, especially their alkali metal or ammonium salts. Examplesinclude: lauroyl sarcosinate, myristoyl sarcosinate, palmitoylsarcosinate, stearoyl sarcosinate oleoyl sarcosinate, or combinationsthereof.

Other suitable anionic surfactants include sodium or potassium alkylsulfates, such as sodium lauryl sulfate, acyl isethionates, acyl methylisethionates, alkyl ether carboxylates, acyl alaninates, acyl gulatames,acyl glycinates, acyl sarconsinates, sodium methyl acyl taurates, sodiumlaureth sulfosuccinates, alpha olefin sulfonates, alkyl benzesulfonates, sodium lauroyl lactylate, sodium laurylglucosideshydroxypropyl sulfonate, and/or combinations.

Zwitterionic or amphoteric surfactants useful herein include derivativesof aliphatic quaternary ammonium, phosphonium, and Sulfonium compounds,in which the aliphatic radicals can be straight chain or branched, andone of the aliphatic substituents contains from 8 to 18 carbon atoms andone contains an anionic water-solubilizing group, e.g., carboxy,sulfonate, sulfate, phosphate or phosphonate. Suitable betainesurfactants are disclosed in U.S. Pat. No. 5,180,577. Typical alkyldimethyl betaines include decyl betaine or2-(N-decyl-N,N-dimethylammonio) acetate, coco-betaine or2-(N-coco-N,N-dimethyl ammonio)acetate, myristyl betaine, palmitylbetaine, lauryl betaine, cetyl betaine, cetyl betaine, stearyl betaine,etc. The amidobetaines can be exemplified by cocoamidoethyl betaine,cocoamidopropyl betaine (CADB), and lauramidopropyl betaine. Othersuitable amphoteric surfactants include betaines, sultaines, sodiumlaurylamphoacetates, alkylamphodiacetates, and/or combinations thereof.

Cationic surfactants useful in the present invention include, forexample, derivatives of quaternary ammonium compounds having one longalkyl chain containing from 8 to 18 carbon atoms such as lauryltrimethylammonium chloride; cetyl pyridinium chloride; cetyltrimethyl-ammonium bromide; cetyl pyridinium fluoride or combinationsthereof.

Nonionic surfactants that can be used in the compositions of the presentinvention include, for example, compounds produced by the condensationof alkylene oxide groups (hydrophilic in nature) with an organichydrophobic compound which may be aliphatic or alkylaromatic in nature.Examples of suitable nonionic surfactants can include the Pluronics®which are poloxamers, polyethylene oxide condensates of alkyl phenols,products derived from the condensation of ethylene oxide with thereaction product of propylene oxide and ethylene diamine, ethylene oxidecondensates of aliphatic alcohols, long chain tertiary amine oxides,long chain tertiary phosphine oxides, long chain dialkyl sulfoxides andcombinations of such materials. Other suitable non-ionic surfactantsincludes alkyl glucamides, alkyl glucosides, and/or combinationsthereof.

The one or more surfactants can also include one or more natural and/ornaturally derived surfactants. Natural surfactants can includesurfactants that are derived from natural products and/or surfactantsthat are minimally or not processed. Natural surfactants can includehydrogenated, non-hydrogenated, or partially hydrogenated vegetableoils, olus oil, passiflora incarnata oil, candelilla cera,coco-caprylate, caprate, dicaprylyl ether, lauryl alcohol, myristylmyristate, dicaprylyl ether, caprylic acid, caprylic ester, octyldecanoate, octyl octanoate, undecane, tridecane, decyl oleate, oleicacid decylester, cetyl palmitate, stearic acid, palmitic acid, glycerylstearate, hydrogenated, non-hydrogenated, or partially hydrogenatedvegetable glycerides, Polyglyceryl-2 dipolyhydroxystearate, cetearylalcohol, sucrose polystearate, glycerin, octadodecanol, hydrolyzed,partially hydrolyzed, or non-hydrolyzed vegetable protein, hydrolyzed,partially hydrolyzed, or non-hydrolyzed wheat protein hydrolysate,polyglyceryl-3 diisostearate, glyceryl oleate, myristyl alcohol, cetylalcohol, sodium cetearyl sulfate, cetearyl alcohol, glyceryl laurate,capric triglyceride, coco-glycerides, lectithin, dicaprylyl ether,xanthan gum, sodium coco-sulfate, ammonium lauryl sulfate, sodium cocoylsulfate, sodium cocoyl glutamate, polyalkylglucosides, such as decylglucoside, cetearyl glucoside, cetyl stearyl polyglucoside,coco-glucoside, and lauryl glucoside, and/or combinations thereof.Natural surfactants can include any of the Natrue ingredients marketedby BASF, such as, for example, CegeSoft®, Cetiol®, Cutina®, Dehymuls®,Emulgade®, Emulgin®, Eutanol®, Gluadin®, Lameform®, LameSoft®, Lanette®,Monomuls®, Myritol®, Plantacare®, Plantaquat®, Platasil®, Rheocare®,Sulfopon®, Texapon®, and/or combinations thereof.

Other specific examples of surfactants include sodium lauryl sulfate,sodium lauryl isethionate, sodium lauroyl methyl isethionate, sodiumcocoyl glutamate, sodium dodecyl benzene sulfonate, alkali metal orammonium salts of lauroyl sarcosinate, myristoyl sarcosinate, palmitoylsarcosinate, stearoyl sarcosinate and oleoyl sarcosinate,polyoxyethylene sorbitan monostearate, isostearate and laurate, sodiumlauryl sulfoacetate, N-lauroyl sarcosine, the sodium, potassium, andethanolamine salts of N-lauroyl, N-myristoyl, or N-palmitoyl sarcosine,polyethylene oxide condensates of alkyl phenols, cocoamidopropylbetaine, lauramidopropyl betaine, palmityl betaine, sodium cocoylglutamate, and the like. Additional surfactants desired include fattyacid salts of glutamate, alkyl glucoside, salts of taurates, betaines,caprylates, and/or mixtures thereof. The oral care composition can alsobe sulfate free.

The oral care composition can comprise one or more surfactants each at alevel from about 0.01% to about 15%, from about 0.3% to about 10%, orfrom about 0.3% to about 2.5%, by weight of the oral care composition.

Thickening Agents

The oral care composition can comprise one or more thickening agents.Thickening agents can be useful in the oral care compositions to providea gelatinous structure that stabilizes the dentifrice and/or toothpasteagainst phase separation. Suitable thickening agents includepolysaccharides, polymers, and/or silica thickeners.

The thickening agent can comprise one or more polysaccharides. Somenon-limiting examples of polysaccharides include starch; glycerite ofstarch; gums such as gum karaya (sterculia gum), gum tragacanth, gumarabic, gum ghatti, gum acacia, xanthan gum, guar gum and cellulose gum;magnesium aluminum silicate (Veegum); carrageenan; sodium alginate;agar-agar; pectin; gelatin; cellulose compounds such as cellulose,microcrystalline cellulose, carboxymethyl cellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxymethyl cellulose,hydroxymethyl carboxypropyl cellulose, methyl cellulose, ethylcellulose, and sulfated cellulose; natural and synthetic clays such ashectorite clays; and mixtures thereof.

Other polysaccharides that are suitable for use herein includecarageenans, gellan gum, locust bean gum, xanthan gum, carbomers,poloxamers, modified cellulose, and mixtures thereof. Carageenan is apolysaccharide derived from seaweed. There are several types ofcarageenan that may be distinguished by their seaweed source and/or bytheir degree of and position of sulfation. The thickening agent cancomprise kappa carageenans, modified kappa carageenans, iotacarageenans, modified iota carageenans, lambda carrageenan, and mixturesthereof. Carageenans suitable for use herein include those commerciallyavailable from the FMC Company under the series designation “Viscarin,”including but not limited to Viscarin TP 329, Viscarin TP 388, andViscarin TP 389.

The thickening agent can comprise one or more polymers. The polymer canbe a polyethylene glycol (PEG), a polyvinylpyrrolidone (PVP),polyacrylic acid, a polymer derived from at least one acrylic acidmonomer, a copolymer of maleic anhydride and methyl vinyl ether, acrosslinked polyacrylic acid polymer, of various weight percentages ofthe oral care composition as well as various ranges of average molecularranges. Alternatively, the oral care composition can be free of,substantially free of, or essentially free of a copolymer of maleicanhydride and methyl vinyl ether.

The thickening agent can comprise one or more inorganic thickeningagents. Some non-limiting examples of suitable inorganic thickeningagents include colloidal magnesium aluminum silicate, silica thickeners.Useful silica thickeners include, for example, include, as anon-limiting example, an amorphous precipitated silica such as ZEODENT®165 silica. Other non-limiting silica thickeners include ZEODENT® 153,163, and 167, and ZEOFREE® 177 and 265 silica products, all availablefrom Evonik Corporation, and AEROSIL® fumed silicas.

The oral care composition can comprise from 0.01% to about 15%, from0.1% to about 10%, from about 0.2% to about 5%, or from about 0.5% toabout 2% of one or more thickening agents.

Prenylated Flavonoids

The oral care composition of the present invention can compriseprenylated flavonoid. Flavonoids are a group of natural substances foundin a wide range of fruits, vegetables, grains, bark, roots, stems,flowers, tea, and wine. Flavonoids can have a variety of beneficialeffects on health, such as antioxidative, anti-inflammatory,antimutagenic, anticarcinogenic, and antibacterial benefits. Prenylatedflavonoids are flavonoids that include at least one prenyl functionalgroup (3-methylbut-2-en-1-yl, as shown in Formula VIII), which has beenpreviously identified to facilitate attachment to cell membranes. Thus,while not wishing to being bound by theory, it is believed that theaddition of a prenyl group, i.e. prenylation, to a flavonoid canincrease the activity of the original flavonoid by increasing thelipophilicity of the parent molecule and improving the penetration ofthe prenylated molecule into the bacterial cell membrane. Increasing thelipophilicity to increase penetration into the cell membrane can be adouble-edged sword because the prenylated flavonoid will tend towardsinsolubility at high Log P values (high lipophilicity). Log P can be animportant indicator of antibacterial efficacy.

As such, the term prenylated flavonoids can include flavonoids foundnaturally with one or more prenyl functional groups, flavonoids with asynthetically added prenyl functional group, and/or prenylatedflavonoids with additional prenyl functional groups synthetically added.

Other suitable functionalities of the parent molecule that improve thestructure-activity relationship (e.g,. structure-MIC relationship) ofthe prenylated molecule include additional heterocycles containingnitrogen or oxygen, alkylamino chains, or alkyl chains substituted ontoone or more of the aromatic rings of the parent flavonoid.

Flavonoids can have a 15-carbon skeleton with at least two phenyl ringsand at least one heterocyclic ring. Some suitable flavonoid backbonescan be shown in Formula IX (flavone backbone), Formula X (isoflavanbackbone), and/or Formula XI (neoflavonoid backbone).

Other suitable subgroups of flavonoids include anthocyanidins,anthoxanthins, flavanones, flavanonols, flavans, isoflavonoids,chalcones and/or combinations thereof.

Prenylated flavonoids can include naturally isolated prenylatedflavonoids or naturally isolated flavonoids that are syntheticallyaltered to add one or more prenyl functional groups through a variety ofsynthetic processes that would be known to a person of ordinary skill inthe art of synthetic organic chemistry.

Other suitable prenylated flavonoids can include Bavachalcone, Bavachin,Bavachinin, Corylifol A, Epimedin A, Epimedin A1, Epimedin B, EpimedinC, Icariin, Icariside I, Icariside II, Icaritin, Isobavachalcone,Isoxanthohumol, Neobavaisoflavone, 6-Prenylnaringenin,8-Prenylnaringenin, Sophoraflavanone G, (−)-Sophoranone, Xanthohumol,Quercetin, Macelignan, Kuraridin, Kurarinone, Kuwanon G, Kuwanon C,Panduratin A, 6-geranylnaringenin, Australone A,6,8-Diprenyleriodictyol, dorsmanin C, dorsmanin F, 8-Prenylkaempferol,7-O-Methylluteone, luteone, 6-prenylgenistein, isowighteone,lupiwighteone, and/or combinations thereof. Other suitable prenylatedflavonoids include cannflavins, such as Cannflavin A, Cannflavin B,and/or Cannflavin C.

Preferably, the prenylated flavonoid has a high probability of having anMIC of less than about 25 ppm for S. aureus, a gram-positive bacterium.Suitable prenylated flavonoids include Bavachin, Bavachinin, CorylifolA, Icaritin, Isoxanthohumol, Neobavaisoflavone, 6-Prenylnaringenin,8-Prenylnaringenin, Sophoraflavanone G, (−)-Sophoranone, Kurarinone,Kuwanon C, Panduratin A, and/or combinations thereof.

Preferably, the prenylated flavonoid has a high probability of having anMIC of less than about 25 ppm for E. coli, a gram-negative bacterium.Suitable prenylated flavonoids include Bavachinin, Isoxanthohumol,8-Prenylnaringenin, Sophoraflavanone G, Kurarinone, Panduratin A, and/orcombinations thereof.

Approximately 1000 prenylated flavonoids have been identified fromplants. According to the number of prenylated flavonoids reportedbefore, prenylated flavonones are the most common subclass andprenylated flavanols is the rarest sub-class. Even though naturalprenylated flavonoids have been detected to have diversely structuralcharacteristics, they have a narrow distribution in plants, which aredifferent to the parent flavonoids as they are present almost in allplants. Most of prenylated flavonoids are found in the followingfamilies, including Cannabaceae, Guttiferae, Leguminosae, Moraceae,Rutaceae and Umbelliferae. Leguminosae and Moraceae, due to theirconsumption as fruits and vegetables, are the most frequentlyinvestigated families and many novel prenylated flavonoids have beenexplored. Humulus lupulus of the Cannabaceae include 8-prenylnaringeninand xanthohumol, which play an important role in the health benefits ofbeer.

The prenylated flavonoid can be incorporated through the hops extract,incorporated in a separately added extract, or added as a separatecomponent of the oral care compositions disclosed herein.

Other Ingredients

The oral care composition can comprise a variety of other ingredients,such as flavoring agents, sweeteners, colorants, preservatives,buffering agents, or other ingredients suitable for use in oral carecompositions, as described below.

Flavoring agents also can be added to the oral care composition.Suitable flavoring agents include oil of wintergreen, oil of peppermint,oil of spearmint, clove bud oil, menthol, anethole, methyl salicylate,eucalyptol, cassia, 1-menthyl acetate, sage, eugenol, parsley oil,oxanone, alpha-irisone, marjoram, lemon, orange, propenyl guaethol,cinnamon, vanillin, ethyl vanillin, heliotropine, 4-cis-heptenal,diacetyl, methyl-para-tert-butyl phenyl acetate, and mixtures thereof.Coolants may also be part of the flavor system. Preferred coolants inthe present compositions are the paramenthan carboxyamide agents such asN-ethyl-p-menthan-3-carboxamide (known commercially as “WS-3”) orN-(Ethoxycarbonylmethyl)-3-p-menthanecarboxamide (known commercially as“WS-5”), and mixtures thereof. A flavor system is generally used in thecompositions at levels of from about 0.001% to about 5%, by weight ofthe oral care composition. These flavoring agents generally comprisemixtures of aldehydes, ketones, esters, phenols, acids, and aliphatic,aromatic and other alcohols.

Sweeteners can be added to the oral care composition to impart apleasing taste to the product. Suitable sweeteners include saccharin (assodium, potassium or calcium saccharin), cyclamate (as a sodium,potassium or calcium salt), acesulfame-K, thaumatin, neohesperidindihydrochalcone, ammoniated glycyrrhizin, dextrose, levulose, sucrose,mannose, sucralose, stevia, and glucose.

Colorants can be added to improve the aesthetic appearance of theproduct. Suitable colorants include without limitation those colorantsapproved by appropriate regulatory bodies such as the FDA and thoselisted in the European Food and Pharmaceutical Directives and includepigments, such as TiO₂, and colors such as FD&C and D&C dyes.

Preservatives also can be added to the oral care compositions to preventbacterial growth. Suitable preservatives approved for use in oralcompositions such as methylparaben, propylparaben, benzoic acid, andsodium benzoate can be added in safe and effective amounts.

Titanium dioxide may also be added to the present composition. Titaniumdioxide is a white powder which adds opacity to the compositions.Titanium dioxide generally comprises from about 0.25% to about 5%, byweight of the oral care composition.

Other ingredients can be used in the oral care composition, such asdesensitizing agents, healing agents, other caries preventative agents,chelating/sequestering agents, vitamins, amino acids, proteins, otheranti-plaque/anti-calculus agents, opacifiers, antibiotics, anti-enzymes,enzymes, pH control agents, oxidizing agents, antioxidants, and thelike.

EXAMPLES

The invention is further illustrated by the following examples, whichare not to be construed in any way as imposing limitations to the scopeof this invention. Various other aspects, modifications, and equivalentsthereof which, after reading the description herein, may suggestthemselves to one of ordinary skill in the art without departing fromthe spirit of the present invention or the scope of the appended claims.

F-Content in the Slurry by ADA One-Minute Release

The fluoride content in the rat caries treatment slurries can beestimated using the ADA One-Minute Release method. The exact methodologyis maintained by the ADA as part of its seal acceptance program and isavailable from the Association. An outline of the method as used hereinis described below. The ADA also references the ANSI/ADA Standard No.116 Oral Rinses or ISO 16408 Dentistry—Oral Hygiene Products—Oral Rinsesfor an example ion-selective electrode method for determining the ionicfluoride content in oral care products.

A stock fluoride solution (0.5 mg/mL, 500 ppm, Ricca, RC3172-16,Arlington, Tex.) was diluted to make solutions of 5 ppm, 25 ppm, 50 ppm,and 250 ppm using deionized water. Each standard was diluted at a 1:1ratio with TISAB II (Ricca, Arlington, Tex.) buffer solution to createcalibration solutions (TISAB IV (Ricca, Arlington, Tex.) if thecalibration solutions are intended for stannous fluoride samples).

A calibration curve was created using the calibration solutions. 200 μLof 25 ppm calibration solution was placed in a microsample cup. Afluoride electrode (VWR, Radnor, Pa.) was placed in the solution and themV of each sample is noted by reading the electrode meter (VWR, Radnor,Pa.). The procedure was repeated for each prepared calibration solution.A calibration curve was constructed by plotting mV vs. Log [F-].

Paste dentifrice samples were prepared by weighing approximately 4 g ofeach paste sample in a 70 mL speed mixer cup. Next, 12 mL of deionizedwas added to the sample cup. The speed mixer cup was mixed for 60seconds (FlackTek Speedmixer, Landrum, S.C.) using Speedmixer Program#7, which is 800 rpm for 5 seconds and then 2200 rpm for 55 seconds. Theslurry samples were transferred into a centrifuge tube and thencentrifuged for 10 minutes at 11,000 rpm.

Samples were prepared for analysis by combining 1 mL of the supernatantand 1 mL of fresh TISAB II buffer for sodium fluoride samples and TISABIV for stannous fluoride samples. The samples were quickly mixed using avortex mixer. 200 μL of the solution were transferred to a micro samplecup. The fluoride electrode was placed in the cap. The value (mV) wasrecorded.

Values for released fluoride were calculated using Formula 1, providedbelow:

ADA Fluoride Released=Average[F]ppm×Dilution Factor×100   Formula 2

[F]=Fluoride Concentration of Sample Supernatant (ppm)

Dilution Factor:

NaF and SnF₂=4−(Formula % Insoluble Raw Materials/100)

*Insoluble Raw Materials (IRM) include silica, titanium dioxide, mica,and prills.

For historical samples for which the slurry fluoride content was notrecorded, the ADA one-minute fluoride release was estimated based on therecorded formulated fluoride content, fluoride source, and abrasivecombination. In some instances, the formulated fluoride content was usedwhen it is known to closely correspond to the ADA one-minute fluoriderelease results, i.e., for Crest® Cavity Protection.

For MFP-containing toothpastes, additional steps were needed to liberatethe fluoride ion such that it could be measured by an ion selectiveelectrode. The steps were the same up to isolation of the supernatant.Following supernatant isolation, a 1.5 mL aliquot of the MFP-containingsupernatant was combined with 2.5 mL of 2N hydrochloric acid (VWR,Radnor, Pa.) in an airtight tube that was caped tightly and mixedvigorously for 30 seconds. The tube headspace was minimized as much aspossible. It was then heated to 50° C. for 20 minutes and cooled to roomtemperature at ambient, benchtop conditions. Once it was cool, 2.575 mLof 2N NaOH (VWR, Radnor, Pa.) was added to the airtight tube, capped,and mixed vigorously for 30 seconds. The resulting solution was allowedagain to cool to room temperature. An aliquot of this was buffered asdescribed above with TISAB II and measured as described above using afluoride ion selective electrode. The appropriate corrections fordilution were made when determining the ADA fluoride released value.

Successful demonstration of an analyst's ability to execute the methodcan be demonstrated by achieving an ADA fluoride released between about95% to 100% for the USP NaF/Silica toothpaste or for about 70% to about85% for the USP SnF₂/Silica toothpaste. The coefficient of correlationfor the F calibration curve on the ion selective electrode should be0.995 or higher. Verify the accuracy by running check standards thatbracket the results before and after sample readings.

Sn-Free In Vitro Plaque Glycolysis and Regrowth Method

The in vitro plaque glycolysis model (iPGRM) is a technique in whichplaque is grown from human saliva and treated with various agents todetermine anti-glycolytic activity of treatments. When bacteria convertsugar into energy with the help of enzymes, acids are formed. Theseacids demineralize and damage the dental enamel. The purpose of thistechnique is to provide a simple method for determining if treatmentcompounds have an inhibitory effect on the metabolic pathways thatplaque microorganisms utilize for the production of acids or toxinsand/or inhibit their growth. For the purposes of the work here, if thetest therapeutic compositions contain Sn, the Sn placebo should betested or controlled for correctly. Additionally, the antibacterialcomposition should be tested with respect to its placebo to determinethe iPGRM value for the antibacterial composition only. This isimportant if buffers, e.g., bicarbonate, orthophosphate, calciumcarbonate, are present in the composition in addition to theantibacterial composition.

A plaque biofilm was grown on glass rods from fresh pooled human salivaand Trypticase Soy Broth (TSB) at 37° C. over 2 days by dipping glassrods into and out of media in a reciprocating motion using a racksuspended on a rod and moved by a reciprocating motor inside of anincubation oven. Treatments were 2 minutes of dentifrice slurry in water(1:5) or diluted treatment in water (1:5). After treatments, biofilmswere incubated with TSB and sucrose until pH indicator showed a colorchange (˜6 hrs). The pH of the media solutions was then measured todetermine the amount of glycolysis inhibition relative to a negativecontrol.

Prior to Day 1, but within a week of Day 1, new glass rods (5 mm×90 mm)were polished approximately 25 mm from the un-tapered end on a lathewith silicon carbide paper of 240, 320, 400, and 600 grit usedsequentially. After the initial polishing, the rods should be polishedwith 600 grit paper before each test. After polishing, rods were storeduntil ready to run. Enough rods should be polished for a full rack oftreatments. A rack can treat 12 compositions with 4 replicates of eachcomposition such that the rack has 48 rods.

On Day 1, saliva was collected daily during the test from a panel of5-10 people by paraffin stimulation and was refrigerated at 4° C. untilit was needed throughout the day. Pool saliva carefully (do not pour inwax/mucus) and mix thoroughly before use. Saliva should be collectedfrom panelists free from disease, who are not on any antibacterialmedication or medication that modifies saliva flow, and who brushregularly with Crest Cavity Protection. The rods were removed fromstorage, rinsed with deionized water to remove any sanding residue,disinfected in 70% ethanol/water solution, and were allowed to dry on asterile surface. Subsequently, the rods were loaded into a hanging rackof holders that were used to dip the rods continuously into media vialscontaining growth media. The rod heights were adjusted and each rod wassecured in place using a rubber o-ring. In the early afternoon, 7 mL ofgrowth media (160 g of a solution of 3% TSB (VWR, Radnor, Pa.) with 3%sucrose (VWR, Radnor, Pa.) was mixed with 240 g pooled human saliva.This TSB/sucrose solution should be sterilized by autoclave beforecombining with the pooled human saliva.) into media vials. The mediavials were arranged under the hanging rods on a rack in an incubationoven. The incubator has been previously modified such that a dippingmotor can dip the rods into the media vials submerging 1.5 cm of the rodinto the growth media at a frequency of 1 dip per minute without therods touching the walls of the media vial. The rods were dippedovernight this way.

On Day 2, an enriched growth media was prepared (500 g of a solution of3% TSB and 10% sucrose was mixed with 33 g pooled human saliva. ThisTSB/sucrose solution should be sterilized by autoclave before combiningwith the pooled human saliva.). This enriched growth media was pipettedinto a new set of media vials (7 mL per vial) and was swapped for theovernight growth media from Day 1. The rods were dipped throughout theday in this enriched growth media for 5 hours at 37° C. in theincubation oven. At the end of the day, a new overnight growth media wasprepared (40 g of a solution of 3% TSB was mixed with 360 g pooled humansaliva and 0.5 g sucrose), pipetted into a new set of media vials, andswapped for the enriched growth media. The rods were dipped overnight inthe same fashion as on the first day.

On Day 3, a glycolysis media was prepared by combining 0.15 g TSB, 25 gsucrose, and 500 mL deionized water resulting in a solution of 0.03% TSBand 0.5% sucrose in water. This solution was mixed then sterilized in anautoclave. The pH was then adjusted to 6.5 using 0.1M HCl and pipettedinto new media vials (7 mL). Two extra vials were filled than wereneeded for the rack of rods as pH blanks. Two drops of chlorophenol redsolution were added to each of the 4 tubes that contained the glycolysismedia for the negative control treatment group (Crest Cavity Protectionslurry). Three drops of bromocresol purple solution were added to 2tubes that contained the glycolysis media for the positive controltreatment group (1% Chlorhexidine solution). Set the rack aside untiltreatments are complete. Two sets of vials were prepared containing 12mL of deionized water to rinse off the treatments. Vials were preparedcontaining the treatment slurries/solutions (7 mL) of homogenizedtreatment and water. The rods were dipped into the treatment vials for 2minutes, rinsed for 10 dips in a first set rinse vials, rinsed for 10dips in a second set of rinse vials, rinsed for 10 dips in a third setof rinse vials, and returned to the incubator rack. It is very importantthat the biofilm on the rods not touch the walls of any of the media,treatment, or rinse vials that would result in the removal of biofilm.The entire biofilm was treated and rinsed. Once all treatments werecomplete, the biofilms on the rods were fully submerged in theglycolysis media inside the incubation oven with no dipping for 2 hours.After two hours, the dipping apparatus was activated. The totalincubation time was between 3 to 7 hours. Incubation is terminated whenthe pH value in the glycolysis media of the negative controls is between4.8-5.6, more ideally 4.9-5.2, and when the pH value in the glycolysismedia of the positive controls is above the negative control. If theindicator dye in the positive control turns yellow, i.e., the pH hasdropped beneath 5.2, the incubation has gone on too long and the testwill need to be repeated.

After incubation termination on Day 3, the rods were removed from theglycolysis media and allowed to dry in the oven. The glycolysis mediawas removed from the incubation oven, allowed to return to roomtemperature, and the pH was measured in each vial and the blank vials todetermine the average pH change of the media following treatment. Thechange in pH is determined with respect to the blank vials. If the finalpH of the blank is less than 6.6, the test needs to be repeated. If thedifference between the positive and negative control is not significantin a student's t-test, the test needs to be repeated. If the change inpH of the negative control of the negative control with respect to theblank is less than 1, the test needs to be repeated.

After the pH values of all the vials were measured, the ΔpH per vial wasdetermined by subtracting its pH from the average pH of the blanks. Theglycolysis inhibition efficacy is determined from the following formula.The average ΔpH of a treatment was determined by averaging the resultsfrom the four replicate vials per treatment.

$\begin{matrix}{{{Efficacy}\mspace{14mu} (\%)} = {100 - {\left( \frac{Avg\Delta pH_{{samp}\; {le}}}{Avg\Delta pH_{{neg}\mspace{11mu} {ctrl}}} \right) \times 100}}} & {{Formula}\mspace{14mu} 3}\end{matrix}$

If the efficacy of the positive control (1% Chlorhexidine solution) isnot between about 65% to about 85% with respect to the negative control(Crest Cavity Protection, Procter & Gamble, Cincinnati, Ohio), the testneeds to be repeated.

In Vitro Plaque Uptake Method for Calcium

The in vitro plaque uptake model (iPUM) is a technique in which plaqueis grown from thawed form frozen human saliva and treated with variousagents to determine the uptake of elemental components into the plaque.A plaque biofilm was grown on glass rods from frozen pooled human salivaand Trypticase Soy Broth (TSB) at 37° over 3 days by dipping glass rodsinto and out of media in a reciprocating motion. Treatments were 2minutes of dentifrice slurry in sterile water (1:5). After treatments,biofilms are dried, weighed, digested and analyzed by either ISE(fluoride) or ICP-OES (Ca, Sn, Zn, etc).

Prior to Day 1, but within a week of Day 1, new glass rods (5 mm×90 mm)were polished approximately 25 mm from the un-tapered end on a lathewith silicon carbide paper of 240, 320, 400, and 600 grit usedsequentially. After the initial polishing, the rods should be polishedwith 600 grit paper before each test. After polishing, rods were storeduntil ready to run test. Enough rods should be polished for a full rackof treatments. A rack can treat 12 compositions with 4 replicates ofeach composition such that the rack has 48 rods. Saliva was collecteddaily from a panel of 5-10 people by paraffin stimulation and frozenovernight. Pool saliva carefully (do not pour in wax/mucus) and mixthoroughly before use. Enough saliva should be frozen prior to the studystarting to last the entire study. Saliva should be collected frompanelists free from disease, who are not on any antibacterial medicationor medication that modifies saliva flow, and who brush regularly withCrest Cavity Protection.

On Day 1, the rods were removed from storage, rinsed with deionizedwater to remove any sanding residue, disinfected in 70% ethanol/watersolution, and were allowed to dry on a sterile surface. Subsequently,the rods were loaded into a hanging rack of holders that were used todip the rods continuously into media vials containing growth media. Therod heights were adjusted and each rod was secured in place using arubber o-ring. In the early afternoon, 7 mL of growth media (160 g of asolution of 3% TSB (VWR, Radnor, Pa.) with 3% sucrose (VWR, Radnor, Pa.)was mixed with 240 g thawed pooled human saliva. This TSB/sucrosesolution should be sterilized by autoclave before combining with thepooled human saliva. Frozen saliva can be thawed in the refrigerator orin a bucket of warm, not hot, water) into media vials. The media vialswere arranged under the hanging rods on a rack in an incubation oven.The incubator has been previously modified such that a dipping motor candip the rods into the media vials submerging 1.5 cm of the rod into thegrowth media at a frequency of 1 dip per minute without the rodstouching the walls of the media vial. The rods were dipped overnightthis way.

On Day 2 and Day 3, an enriched growth media was prepared (500 g of asolution of 3% TSB and 10% sucrose was mixed with 33 g thawed pooledhuman saliva. This TSB/sucrose solution should be sterilized byautoclave before combining with the pooled human saliva.). This enrichedgrowth media was pipetted into a new set of media vials (7 mL per vial)and was swapped for the overnight growth media. The rods were dippedthroughout the day in this enriched growth media for 5 hours at 37° C.in the incubation oven. At the end of the day, a new overnight growthmedia was prepared (40 g of a solution of 3% TSB was mixed with 360 gpooled human saliva and 0.5 g sucrose), pipetted into a new set of mediavials, and swapped for the enriched growth media. The rods were dippedovernight in the same fashion as on the first day.

On Day 4, treatment slurries were prepared by homogenizing 4 g oftreatment composition with 20 g deionized water and pipetted intotreatment vials (7 mL) around such that the rods can be dipped intothem. If the slurries were prepared ahead of time, they should beagitated via pipette immediately prior to treatment to ensure goodsuspension of solid sources of calcium. Two sets of vials were preparedcontaining 12 mL of deionized water to rinse off the treatments. Therods were dipped into the treatment vials for 2 minutes, rinsed for 10dips in a first set rinse vials, rinsed for 10 dips in a second set ofrinse vials, rinsed for 10 dips in a third set of rinse vials, andreturned to the incubator rack. The entire biofilm was treated andrinsed. Once all treatments were complete, the biofilms on the rods werereturned to the incubator oven to dry. It is very important that thebiofilm on the rods not touch the walls of any of the media, treatment,or rinse vials that would result in the removal of biofilm.

The final analysis for calcium occured after the rods were weighed todetermine their weight with biofilm, digesting the rods at 90° C. inconcentrated nitric acid (BDH, Radnor, Pa., Aristar Plus for trace metalanalysis) in a 50 mL conical tube, removing the rods from the digest,cooling and diluting the digest to 50 mL with deionized water, andanalyzing the digest for Ca via inductively coupled plasma-opticalemission spectroscopy (ICP-OES) or inductively coupled plasma-massspectrometry (ICP-MS) (Agilent, Santa Clara, Calif.). Rinsed and driedrods were weighed, and the biofilm mass was calculated by subtractingthe biofilm-free rod mass from the rod mass with biofilm.

The total calcium mass in the biofilm was determined for each rod fromthe analysis of the digest and divided by the biofilm mass for that rodto determine the mass-normalized calcium uptake that was then averagedacross the replicates. The ratio of the calcium uptake between anexperimental treatment and that of Crest Cavity Protection wasdetermined by dividing the mass-normalized calcium uptake for theexperimental treatment averaged across the replicates by themass-normalized calcium uptake for the Crest Cavity Protection treatmentaveraged across the replicates.

The mass-normalized calcium uptake for the Crest Cavity Protection groupshould be about 0.5-3 micro-g_(Ca)/mg_(biofilm). If not, the test shouldbe repeated. The mass-normalized calcium uptake for Tom's of Maine RapidRelief Sensitive (Tom's of Maine, Kennebunk, Me.) should be about 2-3.5times that of Crest Cavity Protection. If not, the test should berepeated.

F-Free HAP Dissolution

The HAP dissolution method was designed to test the acid protection of achosen test dentifrice. After treating hydroxyapatite powder (HAP) withtest dentifrice slurries, the HAP is added to an acidic media and thechange in pH is an indicator of the degree of surface adsorption. Thesmaller the pH rise, the better the surface protection.

Dentifrice slurries (1:3 paste:water) were prepared for the experiment.Specifically, 10 g of dentifrice paste was combined with 30 g ofdeionized water in a 50 mL container with a stir bar. The container wasplaced on a stir plate to mix until the two components were mixed. Thepaste slurries were centrifuged at 15,000 rpm for 15 min to isolatetheir supernatant.

For each treatment, including for the water control, 0.300 g ofhydroxyapatite powder (HAP) was placed into a 50 mL round bottomcentrifuge tube. For treatment with a dentifrice paste, 24 mL of theprepared dentifrice supernatant was added to the HAP. Each treated HAPsample was immediately vortex mixed (DVX-2500 multi-tube vortexer, VWR,Radnor, Pa.) at 2500 rpm for 2 minutes. All samples were thencentrifuged at 15,000 rpm for 15 minutes. The liquid phase was decantedout of the centrifuge tube, which left a HAP pellet. The remaining HAPpellet was rinsed three times by adding deionized water, vortex mixingat 2500 rpm for 1 minute, centrifuging at 15,000 rpm for 15 minutes, andthe liquid phase was decanted out of the centrifuge tube. The treatedHAP pellet was dried in a 55° C. oven overnight.

Samples of HAP were analyzed for ΔpH. 25 mL of 10 mM citric acid (1.9212g of citric acid in 1 L of deionized water) was added to a 50 mL beakerwith a stir bar. The beaker was placed on a stir plate (Metrohm,Herisau, Switerland, Model No. 728) and turned on. The Titrano pHelectrode (Metrohm, Herisau, Switzerland, Model No. 719S) was placed inthe stirring beaker with citric acid. After equilibration of the citricacid solution (until pH has a minimum change of 2.5±0.001 pH within 30second), 50 mg of the dried HAP powder was added to the citric acidsolution. The pH was recorded at 10 min. The % efficacy was determinedby Formula I, below.

$\begin{matrix}{{\% \mspace{14mu} {Efficacy}\mspace{14mu} {{vs}.\mspace{14mu} {Water}}} = \frac{{{{Average}\Delta}\; {pH}\mspace{14mu} {Water}} - {{Average}\mspace{11mu} \Delta \mspace{11mu} {pH}\mspace{14mu} {Treatment}}}{{Average}\mspace{14mu} \Delta \; {pH}\mspace{14mu} {Water}}} & {{Formula}\mspace{14mu} 4}\end{matrix}$

For compositions containing fluoride, the fluoride-free compositionshould be made in order to determine its fluoride-free HAP dissolutionefficacy ensuring that the pH and counter ion content (Na, Sn, etc.) hasbeen properly controlled. For historic samples, the HAP dissolutionefficacy was estimated based on the reported or formulated Sn content ofthe sample using a previously existing relationship. This relationshipis given in FIG. 4.

The method will have been run correctly when the pH change for the watertreated control samples is between about 1.3 and 1.5 pH units at 10minutes. The method should be practiced and repeated until this resultis achieved. For guidance, a pH change for a 1100 ppm F as NaF treatedcontrol sample should be between about 0.9 and 1.1.

Anti-Cavity Performance by Rat Caries, Historical Method

Two rat caries methods have been used in our laboratory over the past 60years to establish anti-caries performance. The first described here isthe Historical Method. The second described later is the Modern Method.The relative performance of actives is similar in the two methods andthe same trends are observed; however, the calculated percent reductionin caries relative to the placebo requires minor adjustment. Thisconversion is achieved by subtracting 5.6% from the Historical Methodscores to give their Modern Method equivalents.

The anti-cavity performance of oral care compositions can besubstantiated in rats using several methods (Stookey, et al. Adv. Dent.Res. 9(3):198-207, 1995). All of the data presented here use methodsdisclosed as part of the FDA Method 37 in the anti-caries monograph. Theexample given here is how our historical data were collected. Thismethod is further described in the following articles: Francis, M D,Arch. Oral Biol. 11:141-1489, 1966; Briner, W W and Francis, M D, CariesRes. 5:180-187, 1971; Donaldson, J D, White, W E, Briner, W W, andCooley, W E, J. Dent. Res. 53:648-652, 1974.

Wistar rats were obtained from Harlan Industries, Inc., Cumberland, Ind.They were shipped via truck and arrived on Day 0 at 22-23 days of age.Animals were received in twenty litters of ten randomly sexed animalsper litter, thus providing the 200 animals used in a standard rat cariesstudy. One animal from each litter was then randomly allocated to one ofthe treatment groups and placed in a numbered stainless-steel,wire-bottom cate (litter mates occupy the same position in each group;e.g., all animals form litter #1 were allocated to the first cage ofeach treatment group). Animals were then weighed in and the weight wasrecorded in the lab book. Animals were fed cariogenic diet #469 adlibitum and deionized water ad libitum.

Treatment required the use of long-stem, cotton-tipped swabs. The swabis dipped into a slurry prepared with toothpaste diluted 1:1 withdeionized water. This dilution was mixed on a stir-plate for fiveminutes prior to treatment application. With the rat's mouth held openby means of a stainless-steel retaining clamp, the dipped swab wasbrushed against the maxillary molars with a front-to-back strokerepeated six times. On the mandible, the swab was dipped into thetreatment slurry and then rotated toward the cheek, thereby movingaround the tongue to reach the mandible molars. Again, six rotations permandible were required. This procedure is repeated on the opposite sideof the mouth with a fresh quantity of toothpaste slurry. Treatment wasapplied twice daily, beginning the Day 1 after arrival, through Day 3.(Day 4, normally a Saturday, and Day 5, normally a Sunday were nottreatment days.) Treatment resumed on Day 6 through Day 10 with theweekend off (Days 11 and 12), and again resumed on Day 13 through Day 17with Days 18 and 19 off. The following week the animals were treated onDay 20 and Day 21, and a final body weight was obtained.

On Day 22, the day after the final treatment day, the animals weresacrificed by decapitation. The tongue was excised and the cheeksincised to the angle of the jaw. A tag with the animal's number (cagenumber) was attached to the snout of the animal with an 8-inch string.The mouth was propped open with a short piece of Tygon tubing. Onceanimals from the entire test were sacrificed, the heads were loweredinto vats of 2% silver nitrate staining solution for one hour. Uponremoval from the stain, the heads were rinsed in at least three changesof running tap water. The heads were then placed in aluminum foil bakingpans, the bottom of the pan covered with tap water and the pan coveredloosely with heavy-gauge aluminum foil.

After staining, the aluminum baking pans containing the heads wereautoclaved at approximately 120° C. and 10 lbs. pressure for 35 minutes,after which the steam was turned off and the pans were allowed to standfor another 15 minutes before the heads were removed. After thisprocedure, the bones containing both the upper and lower molars for eachrat could easily be lifted from the surrounding tissue and placed intothe animal's pre-number plastic vial for future identification. Thesevials were left open for 24-36 hours to dry at room temperature and werethen closed until they were to be sectioned.

Next all the vials for the test were arranged numerically, andmicroscope slides were made up with corresponding animal number (one perrat) and the study number attached. Considering one animal at a time,each quadrant was hemi-sectioned longitudinally, and each section waspermanently mounted on the microscope slide. Each quadrant occupied thesame position on the slide as in the animal's mouth (e.g., the rightupper quadrant was mounted on the right upper corner of the slide).

Using a microscope at 30× magnification, 22 fissures and 24 smoothsurfaces were graded per slide/animal. Each fissure was divided by animaginary line through the middle of its bottom, and then each side ofthe fissure was assigned a severity grade. Since each quadrant wassectioned longitudinally, both halves of each quadrant are graded, andthe most severe grade is recorded for each corresponding smooth surfaceof half fissure. In all there were 68 grades per slide/animal. Themethod of score lesion severity is as follows:

0—no stain in the enamel or dentin at site.

1—dark brown stain in enamel only.

2—dark brown stain in enamel extending to the dentin/enamel junction butno further.

3—stain through the enamel into the dentin.

At the beginning of a study one group of rats was sacrificed to obtain amean zero-time severity score per animal. At the end of the study alltreatment groups were sacrificed and graded to obtain a mean severityscore per animal. Thus in computing the severity of caries in eachparticular group, the 68 smooth surface and half fissure grades for eachanimal in the group were totaled. This number was then divided by thenumber of animals in the group to obtain the mean severity expressed asthe mean number of hypomineralized areas (x HMA).

With all x HMA scores tabulated, a percent reduction is calculated foreach treatment group. This is done by subtracting the score of the testgroup from that of the water or placebo control group and then dividingby the water or placebo control group score. This number is expressed asa percent by multiplying by 100. Further statistical analysis wasperformed to determine significant differences among the groups if theyexisted. A standard analysis of variance was used. Treatments wereranked by a Newman-Keuls analysis. All historical method scores havebeen converted to Modern Method equivalent scores as part ofestablishing the rat caries meta-analysis.

Anti-Cavity Performance by Rat Caries, Modern Method

The test design used here is similar to those found in the FDA Method#37 of the Fluoride Anti-Caries OTC Monograph. The major variations arethe diet used (MIT 200 rather than #469), the caries score method (Keyesmethod rather than HMA), and treatment frequency. Experimentalprocedures were conducted according to the FDA regulations Part 58.

Using litters as a covariate, the use of between 50 and 58 (depending onthe type of fluoride) animals per treatment group satisfies the moststringent power requirements of the ADA's Council on Dental Therapeutics20% clinical difference between treatments at 80% power. However, wehave been routinely using 40 animals per treatment group and both theADA's CDT and the FDA have consistently accepted these tests. Thisrequires initiating the study with 40 animals per group. Twenty-three(23) litters provided these animals. When studies are sized as such,treatment differences of approximately 16% have been found to besignificant on occasion, thus is generally considered the cusp ofclinical significance.

All protocols are reviewed and approved by the Institutional Animal Careand Use Committee prior to the receipt of animals.

The animals were weanling mixed-sex Sprague Dawley rats; weighing 29-53grams. Due to the shipping schedule of the supplier, the dams werereceived with their entire birth litter. The litters were received whenthe pups were 6 days of age and litter size was reduced at 8 days of ageto ten (10) pups per litter. Twenty-five (25) litters were purchased.The five extra litters were to allow for any mortality prior tostratification. Any unused animals were euthanized after the studystratification.

The litters were maintained in large solid-bottom (box-type) cages withdams until the pups were weaned at 18 days of age. Starting at 9 days ofpup age, the dams were rotated daily among the litters until the pupswere weaned at 18 days of age. The pups were maintained in the box cagesuntil 21 days of age. At that time, the pups were stratified and housedin pairs in suspended wire-bottomed cages that had been cleaned andsanitized prior to usage. The change in caging was required to preventartificially increasing the caries rate due to direct contact bedding.The cages were arranged so that all animals of the individual groupswere together and the cages were labeled with group designation andtreatment (treatment code) that the animals received.

When the pups were 21 days of age they were given unique numbers byear-punch with records kept of littermates. Animals were assigned togroups in such a manner that groups were balanced for litter, weight andsex. There were 40 animals per group.

Upon receipt, dams and litters were provided rodent lab diet until thepups were 8 days of age. On day 8 (pup age) dams and litters wereprovided Diet MIT 305. Pups were provided Diet MIT 200 ad libitum at day18 (pup age) and throughout the test period. All animals were providedwith deionized water ad libitum.

Box caging was changed at day 13 and again at day 18 of pup age.Following administration of the inoculum, box cages and the bedding weredecontaminated by autoclaving prior to sanitizing. Cage boards werechanged three times a week at the time when fresh food and water weregiven (Monday, Wednesday and Friday). Clean and sanitized water bottlesand food jars were provided weekly. Suspended caging and banks weresanitized bi-weekly. The animals were observed daily by a staff memberand weekly by the attending veterinarian for any signs of healthproblems. The animals were housed in an AAALAC-accredited facility. Roomtemperature was maintained at 72° F. (±6° F.) with 10-15 air changes perhour and a 12-hour light cycle.

On day 15 (pup age), the animals received an oral inoculation ofstreptomycin-resistant S. sobrinus 6715 (ATCC strain #27352) culture.This involved flooding the mouth with 0.2 ml of culture/animal. On day18 (pup age) the animals were inoculated with S. sobrinus for threeconsecutive days (age 18, 19 and 20 days). This involved placing 0.1 mlof the S. sobrinus culture on the occlusal surfaces of each of themandibular molar quadrants, putting 10 cc of this concentration-adjustedculture into each sanitized and filled water bottle, and lightlyspraying the bedding with no more than 10 cc of the remaining culture.All water bottles were removed and sanitized 24 hours after inoculum hasbeen added. The inoculums were administered to the animals with a 200micropipette.

The treatment phase began at day 22 of pup age. Each treatment had alabeled plastic beaker that was designated for that treatment only.Fresh materials (i.e., obtained from the stock supply) were used foreach treatment. The dentifrices were mixed in a 1:1 ratio (by weight)with deionized water. Specifically, 10 grams of dentifrice was weighedinto a 30 ml beaker; 10 grams of deionized water was then added to thedentifrice. The mixture was then stirred by hand (30 seconds) with aclean micro spatula for the purpose of creating a smooth mixture. Thebeaker containing the slurry and a small magnetic stirring bar wasplaced on a magnetic stirrer, which was set at the lowest speed andallowed to stir for four (4) minutes prior to treatment. The slurry wasprepared immediately prior to each treatment.

A cotton-tipped applicator was dipped into the slurry (for 2 seconds)and was applied to one-half of the rat's mouth in such a way that thesides of the applicator came into contact with both the mandibular andmaxillary molars on one side of the mouth. The treatment wasaccomplished by using a rolling motion of the sides of the applicatorover the mandibular and maxillary molar teeth for 15 seconds. Theapplicator was dipped into the slurry for the second time (again, for 2seconds) and the other side of the rat's mouth similarly treated for 15seconds. A new applicator was used for each animal.

Treatments were administered twice daily for five days with a singledaily treatment on weekends. The first treatment each day began atapproximately the same time every day, and the second treatment didbegin no earlier than six hours after the first treatment. Singulartreatments were given at a 24-hour interval on weekends. Treatmentmaterials were stored at room temperature. All treatment products werereturned to sponsor at study completion.

One week after the initiation of the inoculation regimen and at studytermination, an oral swabbing was taken from each rat using a sterilecotton swab (six-inch, single-tipped applicator). The microorganisms onthe mandibular and maxillary molar teeth were sampled, using a rollingmotion of the swab for 15 seconds on one side of the mouth, rolled overthe tongue, and rolled over the molar teeth on the other side of themouth for an additional 15 seconds. Immediately after the applicator wasremoved from the animal's mouth, it was streaked across half of a 100 mmpetri plate containing Mitis Salivarius agar to which 200 units/ml ofstreptomycin sulfate was added. The plates were incubated for 48 hoursat 37° C. with 10% CO₂. The colony count taken after the 48 hours ofincubation was recorded in the logbook.

The experimental duration of the rat caries studies is three weeks.Immediately prior to termination, all animals were observed for anyvisual signs of ill health or pathology, individually weighed and anoral swabbing taken to confirm S. sobrinus implantation. The animalswere euthanized by carbon dioxide inhalation. Code numbers were assignedto each animal and the heads were then removed, placed in individualjars along with the code number, and cooked under pressure (10 PSI for12 minutes). The hemijaws were then removed and freed of all softtissue.

The cleaned hemijaws (four quadrants) were put into plastic vials withthe code numbers taped to the vial. A murexide solution (0.3 g murexide;300 ml DI H20 and 700 ml of ethanol) was added to each vial and the jawswere allowed to stain overnight. The jaws were then rinsed and allowedto air dry.

The hemijaws were microscopically examined for smooth surface caries,sectioned, and then microscopically examined for sulcal andinterproximal caries using the Keyes Method. The scoring method isdetailed in Navia, J N, Animal Models in Dental Research, pp 287-290,1977; and Keyes, P H, J. Dent. Res. 37:1088-1099, 1958. All analyseswere performed using SAS statistical software, version 9.4. The groupswere compared using analysis of variance (ANOVA), with a fixed effectfor group and a random effect for litter. The litter effect was includedin the models to reduce a known factor affecting the variability of themeasurements. Pair-wise comparisons between groups were made usingTukey's multiple comparisons procedure to control the overallsignificance level (α=0.05) of the comparisons.

The specific types of data, which were tabulated, and statisticallyanalyzed may include:

-   -   1) Mortality Data Experimental Phase        -   a. Initial number of animals        -   b. Final number of animals        -   c. Percent mortality    -   2) Growth Data Experimental Phase        -   a. Initial body weight (mean±S.E.M.)        -   b. Final body weight (mean±S.E.M.)    -   3) Caries Experience        -   a. Enamel and dentinal involvement of smooth surface            (buccal, lingual) lesions (mean±S.E.M.)        -   b. Enamel and dentinal involvement of interproximal lesions            (mean±S.E.M.)        -   c. Enamel and dentinal involvement of total smooth surface            (buccal, lingual & interproximal) lesions (mean±S.E.M.)        -   d. Enamel and dentinal involvement of sulcal lesions            (mean±S.E.M.)        -   e. Total caries involvement combining the scores from the            Keyes method of scoring smooth surface, interproximal, and            sulcal caries (mean±S.E.M.)

Preparation of Oral Care Compositions

The oral care compositions of TABLE 1A were prepared by combining one ormore humectants, water, sweetener(s), tin ion source, sodium gluconate,and/or flavor(s) to create a liquid mixture. The liquid mixture washomogenized at 25° C. for 2 minutes. Next, sodium hydroxide (50%solution) was added to the liquid mixture and the liquid mixture washomogenized at 25° C. for 2 minutes. A separate powder mixture wasprepared by combining a portion of the calcium ion source and anythickening agents, such as xanthan gum and/or sodiumcarboxymethylcellulose. The powder mixture was then combined with theliquid mixture. Next, the surfactant, such as sodium lauryl sulfate, wasadded to the mixture. The contents were homogenized at 25° C. for 2minutes. The hops extract was then combined with the mixture andhomogenized at 25° C. for 2 minutes. Finally, the remaining portion ofthe calcium ion source and the buffering agent were combined with themixture and homogenized at 25° C. for 2 minutes.

Preparation of Commercial Oral Care Compositions with Hops Beta Acid

The commercial oral care compositions were combined with hops beta acidby weighing out a portion of commercial oral care composition and mixingin the appropriate amount of hops beta extract. The combined oral carecomposition was homogenized at 25° C. for at least 2 minutes.

Ex. 1a, 1b, 1c, and 1d, as shown in TABLE 1A, were prepared inaccordance with the Experimental Methods, described above. The Hops BetaAcids were supplied by Hopsteiner® as an extract from Humulus lupulus.The Hopsteiner® extract was approximately 45%, by weight of the extract,of hops beta acids and less than 1%, by weight of the extract, of hopsalpha acids. Ex. 1a and 1c have 13% water while Ex. 1b and 1d do nothave any separately added water. Ex. 1a and Ex. 1b are free fromfluoride while Ex. 1c and 1d have sodium monofluorophosphate.

TABLE 1A Compositions Ex. 1a Ex. 1b Ex. 1c Ex. 1d (wt %) (wt %) (wt %)(wt %) Sorbitol 37.98  — 36.83  — Glycerin — 49.00  — 47.70  Water13.00  — 13.00  — Sodium Gluconate 1.00 1.00 1.00 1.00 Stannous Chloride1.10 1.10 1.10 1.10 Calcium Carbonate 32.00  32.00  32.00  32.00 Xanthan Gum 0.30 0.50 0.30 0.50 Carboxymethyl 1.00 — 1.00 — CelluloseCarbomer — 1.00 — 1.00 Sodium Lauryl 1.29 1.40 1.29 1.40 Sulfate Flavor1.00 1.10 1.00 1.10 Sodium Saccharin 0.50 0.40 0.50 0.40 SteviaGlycosides — 0.30 — 0.30 Sodium Hydroxide 0.33 1.20 0.33 1.35 Hops BetaAcid 0.50 0.50 0.50 0.50 Extract* Sodium 10.00  10.00  10.00  10.00 Bicarbonate Titanium Dioxide — 0.50 — 0.50 Sodium — — 1.15 1.15Monofluorophosphate *Hops Beta Acid Extract supplied by Hopsteiner ®,with 45% hops beta acids and less than 1% hops alpha acids

TABLE 1B Hops Beta Acids Extract Specification Ingredient Amount (wt %)Hops Beta Acids 45 ± 2  Hops Alpha Acids 0.4 ± 0.3 Hops oils 1.5 ± 0.5Propylene Glycol 20 ± 15 Water <8% pH  11 ± 0.5

TABLE 1B describes the hops beta acid extract provided by Hopsteiner®.Since the hops beta acids are provided as an extract, there can be somevariability in the amounts of certain ingredients. However, the extractcomprises approximately 45%, by weight of the extract, of the hops betaacids and approximately 0.4%, by weight of the extract, of hops alphaacids. This is dramatically different to previous hops extracts whichtypically have more hops alpha acids than hops beta acids. Other minoringredients may be present in the Hops Beta Acid extract.

TABLE 2 Performance of Compositions % % Total Reduction ReductionTreatment Leg Caries v Placebo Predicted ADA iPGRM iPUM-Ca HAP SilicaPlacebo 42.36 0 0 1 0 Silica 1000 ppm 34.08* 20 20 1000 0 1 0 F as MFPSilica 1100 ppm 27.39* 35 39 1100 0 1 0 F as NaF Ex. 1b 27.78* 34 38 057 3.6 33 Ex. 1d 21.43* 50 65 1100 57 3.9 33 *Significantly differentfrom Silica Placebo, p < 0.05 as determined by Tukey's adjustment formultiple comparisons

TABLE 3 Fluoride Vector % Reduction % Reduction Performance TreatmentLeg v Placebo Predicted ADA Threshold 2800 ppm F as 62 63 2800 ADA >650ppm NaF Dentifrice F as NaF in a 1450 ppm F as 49 45 1450 dentifrice NaFDentifrice results in a 29%. 1100 ppm F as 47 39 1100 reduction in ratNaF Dentifrice caries following 650 ppm F as 24 29 650 meta-analysis NaFDentifrice 500 ppm F as 25 500 NaF Dentifrice 250 ppm F as 14 17 250 NaFDentifrice Placebo — — 0 Dentifrice

TABLE 4 Antibacterial Agent Vector % Reduction % Reduction Treatment Legv Water Predicted iPGRM 0.5% Chlorhexidine 7.5 8 75 Solution 0.2%Chlorhexidine 3.8 4.2 49 Solution Water — — 1.0% Chlorhexidine 27   11.196 Solution 0.1% Chlorhexidine 2.4 1.3 29 Solution Water — —

TABLE 5 Calcium Vector % Reduction % Reduction Treatment Leg v WaterPredicted iPUM-Ca 1.75% CaCl₂ 5.6 10 4.2 Composition 2.5% Ca 1.2 5.4 3.1Glycerophosphate Composition Water — —

TABLE 6 HAP Dissolution Reduction Vector % Reduction % ReductionTreatment Leg v Control Predicted HAP 1000 ppm F as SnF₂   3.4   9.0 13Dentifrice + 9000 ppm Sn 1000 ppm F as SnF₂ — — 0 Dentifrice 1100 ppm Fas SnF₂ 19 10 14 Dentifrice + 10,000 ppm Sn 1100 ppm F as SnF₂ 14 10 14Dentifrice + 10,000 ppm Sn 1100 ppm F as SnF₂ 12 12 17 Dentifrice +12,500 ppm Sn 1100 ppm F as NaF — — 0 Dentifrice

TABLE 7 Arginine Bicarbonate/Calcium Carbonate Example % Reduction %Reduction Treatment Leg v Control Predicted Sig ADA iPGRM iPUM-Ca HAP1100 ppm F as 22 39 s 1100 0 1 0 NaF/Silica Silica F-Placebo — — —Toothpaste Arginine Bicarb/ −8 6.3 ns 0 9 3 0 Calcium Carb

The rat caries results, presented in FIG. 1/TABLE 1, indicated that Ex1b reduced caries relative to sodium fluoride/silica andmonofluorophosphate/silica compositions in a placebo-controlled study.Ex. 1b which did not have fluoride, had a total caries reductionequivalent to the sodium fluoride/silica control product; however, ithad an unexpectedly large reduction in sulcal caries relative tocontrol. The disclosed compositions were designed to work on the toothand in the biofilm with a collection of subtherapeutic compositions.This has led to an unexpected reduction in sulcal and interproximalcaries with respect to the control compositions. When fluoride is addedto the disclosed compositions, as in Ex. 1d, the reduction in caries isenhanced with a notable reduction in smooth surface caries.

It was unexpectedly found that the non-fluoride mechanisms can have alarge contribution to the reduction in caries considering that such anapproach had not be disclosed in the art. In order to better understandthis result, a set of laboratory tests were performed to measure thecontribution of each subtherapeutic composition to the compositiontested in the rat. Individual methods have been developed to measure theefficacy of compositions along the above described intervention vectors.These methods are the: i) Sn-Free in vitro plaque glycolysis andregrowth method (Sn-Free iPGRM); ii) in vitro plaque uptake method forcalcium (iPUM-Ca); iii) F-Free hydroxyapatite solubility reductionmethod (F-Free HAP); and iv) ADA one-minute fluoride release (ADA). Aperson of ordinary skill in the art would recognize that someingredients, such as Sn, have both an antibacterial effect and a HAPsurface stabilization effect. Such behavior complicates the analysis ofa composition's performance; therefore, the entire contribution of suchingredients is considered through a single mechanism only. Thus, for thepurposes of this invention, a composition's antibacterial efficacyshould be determined with respect to its Sn placebo. Similarly, for thepurposes of this invention, a composition's ability to reducehydroxyapatite solubility should be determined using the fluoride-freeversion of the composition (if it contains fluoride). TABLE 2illustrated the results of the characterization of the novelcompositions and control toothpastes using the four different methodsindicated above with their corresponding rat caries scores, as disclosedin the Example section.

During the discovery of fluoride, several rat caries tests were runexploring both the effectiveness of fluoride as well as alternatives.The catalogue of rat caries experiments were analyzed to find exampleswhere the use of a single anticaries agent contributed an anticarieseffect.

TABLE 3 shows the variation in anticaries efficacy with respect tosoluble fluoride content as determined by the ADA method. Using thisexample and the meta-analysis, a 650 ppm fluoride as sodium fluoride wasestimated to effect a reduction in caries of about 29%, or about 25%, orabout 30%, with respect to the placebo or water control in rat cariesexperiments. The table indicates the expected caries reduction in therat model for the transition from subtherapeutic to therapeutic levelsof fluoride delivered as sodium fluoride.

TABLE 4 shows the variation in anticaries efficacy with respect toSn-Free antibacterial activity as measured by the Sn-Free iPGRM. Sincethe iPGRM measures reduction in antibacterial efficacy through changesin the pH, non-antibacterial agents that modify pH, such as sodiumbicarbonate, can be corrected for when determining the efficacy of theantibacterial agent. This can be done using placebo controls asnecessary. The contribution of Sn was measured through the solubilityreduction and is excluded from this test. The iPGRM dose response ofchlorhexidine is shown in FIG. 2.

TABLE 5 shows the variation in anticaries efficacy with respect tocalcium co-ion effect as measured by the iPUM. This method measures thetotal calcium uptake of both soluble and insoluble sources. If aninsoluble source was used, calcium sources were preferred that weresubstantially more soluble than hydroxyapatite so that they willdissolve preferentially when exposed to plaque acids. For example,dicalcium phosphate or calcium carbonate dissolve readily on exposure toacid while calcium pyrophosphate is a poor source of calcium for thepurposes here. Calcium pyrophosphate does not readily dissolve onexposure to acid and is a poor source of calcium for the purposes ofprotecting teeth from plaque acids through a co-ion effect.Nevertheless, insoluble sources have the advantage of increasedresidence time in the plaque and can, therefore, provide a bloom ofcalcium to correspond temporally to the generation of plaque acid. TheiPUM dose response of calcium-containing compositions is shown in FIG.3.

TABLE 6 shows the variation in anticaries efficacy with respect to theF-Free reduction in HAP solubility as measured by F-free HAP. Thecontribution of the fluoride co-ion effect is measured through the ADAmethod and is excluded from this test. An example of the F-free HAPresponse for different levels of Sn in Sn/silica toothpastes is given inFIG. 4.

Finally, the threshold can be defined for which combinations ofsubtherapeutic compositions result in a therapeutic composition. Onaverage, this transition can be defined for those compositions providinga reduction in rat caries of approximately 29% with a statisticallysignificant reduction in caries with respect to the water orF-placebo/silica toothpaste negative control. Of course, performance canbe substantiated using an actual rat caries performance test.

With these methods, the efficacy can be quantified of variouscompositions along the important anticavity intervention methods. Thesame analysis can be retrospectively applied to a long history of ratcaries experiments conducted during the original development offluoride-containing anticavity compositions. Approximately 170 ratcaries experiments were analyzed containing over 800 individualtreatments for the reductions of caries in two different, but similar,rat caries models were analyzed. These experiments were conductedbetween 1959 and 2019. Rat caries is the preferred animal model forhuman caries and is included in the US Anticaries Monograph (21 CFR part355) to ensure the efficacy of fluoride-containing products. It isadditionally sensitive to non-fluoride anticaries mechanisms andcompositions. Finally, animal models for caries are generally consideredinterchangeable as long as they have been properly developed.

This retrospective analysis can help define the scope of the presentinvention. As described herein, the present invention is directed to acombination of subtherapeutic compositions can result in therapeuticcomposition when tested in total. The examples can allow for the settingof performance thresholds for the subtherapeutic to therapeutictransition in each mechanism as defined by the specified performancetest. However, those results, alone, might not anticipate the combinedperformance of the compositions in a rat caries experiment. Thus, thethresholds for the transition from a subtherapeutic to a therapeuticdose for each mechanism were modeled with historical rat caries data.The analysis gave a model from which can predict reductions in ratcaries for combinations of subtherapeutic compositions as describedherein. In combination, the examples, performance thresholds, andmathematical model can help define the scope of the present invention.

The retrospective analysis of rat caries experiments was possiblebecause of detailed descriptions of the treatments/ingredients withadditional measurements of F content, Sn content, and pH frequentlydocumented in the experimental record. Efficacy measures in the variousanticaries mechanism methods described above were either measureddirectly when the ingredients could be obtained or estimated using thedetailed description of the treatments and comparisons to similarpresent-day compositions. Estimations of rat caries efficacy using thefour methods named above resulted in a model correlation coefficient,r², of ˜0.76. The correlation coefficient suggests that 76% of thevariation in rat caries efficacy is captured by these methods. Theremaining 24% of variation is typically ascribed to biological variationtypical in biological methods. Suffice it to say, we believe thisrepresents a good model for the performance of various compositions inrat caries.

The prediction formula is reproduced below for the % reduction in ratcaries with respect to water or a silica-based-abrasive,placebo-toothpaste negative control.

% Reduction=1.77*sqrt(ADA/2)+0.146*iPGRM+3.97*iPUM-Ca+0.689*HAP−6.84  Formula 1

The predicted versus actual values for the retrospective analysis of ratcaries data are given in FIG. 5a with a plot of the residuals in FIG. 5b. Additionally, 16 is subtracted from the % Reduction calculated usingFormula 1 if a MFP fluoride source is used. This was done because MFP isless effective in the rat caries model.

The dimensions and values disclosed herein are not to be understood asbeing strictly limited to the exact numerical values recited. Instead,unless otherwise specified, each such dimension is intended to mean boththe recited value and a functionally equivalent range surrounding thatvalue. For example, a dimension disclosed as “40 mm” is intended to mean“about 40 mm.”

Every document cited herein, including any cross referenced or relatedpatent or application and any patent application or patent to which thisapplication claims priority or benefit thereof, is hereby incorporatedherein by reference in its entirety unless expressly excluded orotherwise limited. The citation of any document is not an admission thatit is prior art with respect to any invention disclosed or claimedherein or that it alone, or in any combination with any other referenceor references, teaches, suggests or discloses any such invention.Further, to the extent that any meaning or definition of a term in thisdocument conflicts with any meaning or definition of the same term in adocument incorporated by reference, the meaning or definition assignedto that term in this document shall govern.

While particular embodiments of the present invention have beenillustrated and described, it would be obvious to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the invention. It is thereforeintended to cover in the appended claims all such changes andmodifications that are within the scope of this invention.

What is claimed is:
 1. An oral care composition comprising: (a) a firstsubtherapeutic anticaries agent; (b) a second subtherapeutic anticariesagent, wherein the oral care composition is free of fluoride and thefirst and second subtherapeutic agents collectively have a therapeuticanticavity benefit.
 2. The oral care composition of claim 1, wherein thefirst subtherapeutic anticaries agent comprises metal ion source.
 3. Theoral care composition of claim 2, wherein the metal ion source comprisestin, zinc, or combinations thereof.
 4. The oral care composition ofclaim 3, wherein the tin comprises stannous chloride.
 5. The oral carecomposition of claim 3, wherein the zinc comprises zinc phosphate, zinclactate, zinc oxide, zinc citrate, or combinations thereof.
 6. The oralcare composition of claim 1, wherein the oral care composition comprisesamino acid, the amino acid comprising glycine, alanine, valine,isoleucine, tryptophan, phenylalanine, proline, methionine, leucine,serine, threonine, tyrosine, asparagine, glutamine, cysteine,citrulline, aspartic acid, glutamic acid, lysine, arginine, histidine,or combinations thereof.
 7. The oral care composition of claim 1,wherein the second subtherapeutic anticaries agent comprises hops acid,sugar alcohol, urea, or combinations thereof.
 8. The oral carecomposition of claim 7, wherein the hops acid comprises hops beta acid.9. The oral care composition of claim 1, wherein the therapeuticanticavity benefit comprises the anticavity benefit provided by acontrol composition, the control composition comprising at least 650 ppmof free fluoride ions.
 10. The oral care composition of claim 1, whereinthe therapeutic anticavity benefit comprises a rat caries score equal toor greater than the rat caries score of a control composition comprisingat least 650 ppm of free fluoride ions.
 11. The oral care composition ofclaim 1, wherein the therapeutic anticavity benefit comprises areduction in greater than 25% of cavities in a rat caries test.
 12. Theoral care composition of claim 1, wherein the therapeutic anticavitybenefit comprises a reduction in at least about 29% of cavities in a ratcaries test.
 13. An anticavity, fluoride-free oral care composition,wherein the composition has a rat caries score of about 60% or more of arat caries score of a positive control oral care composition, thepositive control oral care composition comprising 1100 ppm of sodiumfluoride.
 14. The anticavity, fluoride-free oral care composition ofclaim 13, wherein the oral care composition comprises hops acid, aminoacid, sugar alcohol, metal ion source, or combinations thereof.
 15. Theanticavity, fluoride-free oral care composition of claim 13, thecomposition comprising at least two of: (a) first antibacterial agent;(b) second antibacterial agent; and/or (c) calcium.
 16. The anticavity,fluoride-free oral care composition of claim 13, the compositioncomprising: (a) antibacterial agent; (b) buffering agent; and (c)calcium.
 17. The anticavity, fluoride-free oral care composition ofclaim 16, wherein the antibacterial agent comprises hops acid, sugaralcohol, or combinations thereof.
 18. The anticavity, fluoride-free oralcare composition of claim 17, wherein the antibacterial agent compriseshops beta acid.
 19. The anticavity, fluoride-free oral care compositionof claim 16, wherein the antibacterial agent comprises metal ion source.20. The anticavity, fluoride-free oral care composition of claim 19,wherein the metal ion source comprises tin, zinc, or combinationsthereof.
 21. The anticavity, fluoride-free oral care composition ofclaim 20, wherein the tin comprises stannous chloride.
 22. Theanticavity, fluoride-free oral care composition of claim 13, wherein theoral care composition comprising amino acid, the amino acid comprisesglycine, alanine, valine, isoleucine, tryptophan, phenylalanine,proline, methionine, leucine, serine, threonine, tyrosine, asparagine,glutamine, cysteine, citrulline, aspartic acid, glutamic acid, lysine,arginine, histidine, or combinations thereof.
 23. The anticavity,fluoride-free oral care composition of claim 16, wherein the calciumcomprises calcium carbonate, calcium bicarbonate, calcium phosphate,calcium chloride, or combinations thereof.
 24. An oral care compositioncomprising: (a) a therapeutic anticaries agent comprising fluoride; and(b) a subtherapeutic anticaries agent, wherein the subtherapeuticanticaries agent is free of fluoride, wherein the oral care compositionhas a therapeutic benefit greater than the therapeutic anticaries agent.